Reporting Checklist for Nature Neuroscience

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1 Corresponding Author: Manuscript Number: Manuscript Type: George Mentis NNA57012C Article Reporting Checklist for Nature Neuroscience # Main s: 8 # Supplementary s: 16 # Supplementary Tables: 3 # Supplementary Videos: 0 This checklist is us to ensure good ing standards and to improve the reproducibility of publish results. For more information, please read Reporting Life Sciences Research. Please e that in the event of publication, it is mandatory that authors include all relevant methodological and statistical information in the manuscript. Statistics ing, by figure example example Please specify the following information for each panel ing quantitative data, and where each item is (section, e.g. Results, & paragraph number). Each figure should ideally contain an exact sample size (n) for each experimental group/condition, where n is an exact number and a range, a clear definition of how n is defin (for example x cells from x slices from x animals from x litters, collect over x days), a description of the statistical us, the results of the s, any descriptive statistics and clearly defin error bars if applicable. For any experiments using custom statistics, please indicate the us and stats obtain for each experiment. Each figure should include a statement of how many times the experiment shown was replicat in the lab; the details of sample collection should be sufficiently clear so that the replicability of the experiment is obvious to the reader. For experiments in the text but in the figures, please use the paragraph number instead of the figure number. e: Mean and standard deviation are appropriate on small samples, and plotting independent data points is usually more informative. When technical replicates are, error and significance measures reflect the experimental variability and the variability of the biological process; it is misleading to state this clearly. FIGURE NUMBER 1a results, para 6 TEST USED WHICH TEST? oneway ANOVA unpair t SECTION & PARAGRAPH # Results para 6 EXACT VALUE 9, 9, 10, 15 n DEFINED? from at least 3 litters/group 15 slices from 10 SECTION & PARAGRAPH # Methods para 8 Results para 6 DESCRIPTIVE STATS (AVERAGE, VARIANCE) REPORTED? mean / SEM mean / SEM SECTION & PARAGRAPH # Results para 6 P VALUE EXACT VALUE p = p = SECTION & PARAGRAPH # Results para 6 DEGREES OF FREEDOM & F/t/z/R/ETC VALUE VALUE F(3, 36) = 2.97 t(28) = SECTION & PARAGRAPH # Results para 6 1

2 FIGURE NUMBER 1c 1d 1e 1g TEST USED WHICH TEST? Oneway Oneway Oneway unpair t ; Mann SECTION & PARAGRAPH # EXACT VALUE n = 8, 6, 12 n = 8, 6, 12 n = 8, 6, 12 n = 17, 13 n DEFINED? P2 L2 motor and SMA (INPUT RESISTANCE) P2 L2 motor and SMA (TIME CONSTANT) P2 L2 motor and SMA (RHEOBASE) P2 L2 motor neurons from 3 per WT and SMA group (somatic & dendritic synapses) SECTION & PARAGRAPH # DESCRIPTIVE STATS (AVERAGE, VARIANCE) REPORTED? SECTION & PARAGRAPH # P VALUE EXACT VALUE P < (post hoc: wt unaffect P=0.999; wt affect smaaffect unaffect P<) P = (post hoc: wt unaffect P=0.977; wt affect P=0.0030; smaunaffect v smaaffect P=0.0052) P = unaffect P=0.942; wt v smaaffect P=0.0046; smaunaffect v smaaffect P=0.0059) t WT vs SMA soma P = t WT vs SMA dendritic density 050 μm P = 0.09; Mann μm P = 0.24; μm P = SECTION & PARAGRAPH # & text & text & text DEGREES OF FREEDOM & F/t/z/R/ETC VALUE VALUE F (2, 23) = F (2, 23) = F (2, 23) = Soma t = df = 28 dendritic density 050 μm t = df = μm U = μm U = 41 SECTION & PARAGRAPH # 2

3 1i 2b Oneway Oneway n=11, 6, 4 n=11, 6, 6, 8 P2 L2 motor and SMA (EPSP), SMA, SMAPV Cre, SMAChAT Cre SMA P = 0.03 unaffect P=0.9516; wt affect P=0.0265; smaunaffect v smaaffect P=0.0734) Input resistance: P = ; P=0.0051; P=0.9982; wt P=0.0031; P=0.0118; sma P=0.988; P=0.0092) Time constant: P = ; P=0.0298; P=0.8848; wt P=0.0159; P=0.1460; sma P=0.9982; P=0.1175) Capacitance: P = P=0.9832; P=0.9559; wt P=0.9754; P=0.9994; sma P=0.8913; P=0.8249) F (2, 18) = Input resistance: F (3, 27) = 6.228; Time constant: F (3, 27) = 5.165; Capacitance: F (3, 27) =

4 2d 2g Oneway Oneway n=11, 6, 6, 8 n=4, 4, 4, 5, SMA, SMAPV Cre, SMAChAT Cre (EPSP), SMA, SMAPV Cre, SMAChAT Cre P = P=0.0006; P=0.2912; wt P=0.0060; P=0.1386; sma P=0.7107; P=0.5499) WT v SMA P=0.008; WT v SMA PvCre P= WT v SMA ChATCre P= SMA v SMA PvCre P= SMA v SMA ChATcre P= SMA SMAChATCre P= F (3, 27) = F (3, 13) =

5 2i,j Oneway (i): n=17,14, 13,15 (j): n=62, 46, 43, 26 neurons from 3 per WT and SMA group (i: somatic and j: dendritic) (i): P< P=0.2404; wt P=0.0025; P=0.0208; sma P=0.4841; P=0.3667) (j): WT vs SMA dendritic density 050 μm P < P=0.5677; wt P=0.0010; P=0.0087; sma P=0.8438; P=0.0929) WT vs SMA dendritic density μm P = P=0.0007; P=0.2669; wt P=0.0052; P=0.1043; sma P=0.9672; P=0.3042) (i): (3,55) = (j):wt vs SMA dendritic density 050 μm F (3, 190) = WT vs SMA dendritic density μm F (3, 113) =

6 3b P2 data Mann P4 and P11 data Oneway n = 3, 4, 3, 4, 4 P2, P4, P11, L2 motor neurons from 3 WT, 4 SMA, 3 SMAPV Cre, 4 SMAChAT Cre and 4 SMA(PVChAT) Cre P2 WT vs SMA P = P4 P = P=0.0008; P=0.0127; wt P=0.1341; wt v (Pv Chat)Cre P=0.2195; P=0.6831; sma P=0.0478; sma v (Pv Chat)Cre P=0.0262; P=0.5129; (Pv Chat)Cre P=0.3502; v (Pv Chat)Cre P=0.9982) P11 P = P=0.0011; P=0.0166; wt P=0.8114; wt v (Pv Chat)Cre P=0.3385; P=0.7525; sma P=0.0079; sma v (Pv Chat)Cre P=0.0423; P=0.1045; (Pv Chat)Cre P=0.3835; v (Pv Chat)Cre P=0.8992) P2 WT vs SMA U = 3.5 P4 F (4, 13) = P11 F (4, 12) =

7 4b 4d Oneway unpair t P2 L2 motor n = 7, 6, 4 and SMA n = 8, 6 and SMA 10 pa current WT vs SMA P = pa P = pa P = pa P = pa P = pa P = pa P = pa P = pa P = pa current WT vs SMA P = pa P = pa P = pa P = pa P = pa P = pa P = pa P = pa P = pa F (2, 14) = pa F (2, 14) = pa F (2, 14) = pa F (2, 14) = pa F (2, 14) = pa F (2, 14) = pa F (2, 14) = pa F (2, 14) = pa F (2, 14) = pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t =

8 4e 4f 5b unpair t unpair t Oneway n = 6, 6 n = 6, 7 n = 4, 3, 3, 3, 3 neurons from SMA PV Cre and SMA neurons from SMA ChAT Cre and SMA P4 QL NMJs from 4 WT, 3 SMA, 3 SMA PV Cre, 3 SMA ChAT Cre and 3 SMAPVChAT Cre 10 pa current PV Cre SMA vs SMA P = pa P = pa P = pa P = pa P = pa P = pa P = pa P = pa P = pa current ChAT Cre SMA vs SMA P = pa P = pa P = pa P = pa P = pa P = pa P = pa P = pa P = 0.32 P < 10 pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t = pa t = F (4, 11) =

9 5d 5f 5g Oneway Oneway unpair t n = 4, 4, 3, 3, 3 n = 4, 4, 3, 3, 3 n = 10, 7 P4 QL CMAP responses from 4 WT, 4 SMA, 3 SMA PV Cre, 3 SMA ChAT Cre and 3 SMAPVChAT Cre (AMPLITUDE OF CMAP) P4 QL CMAP responses from 4 WT, 4 SMA, 3 SMA PV Cre, 3 SMA ChAT Cre and 3 SMAPVChAT Cre Righting reflex times from 10 SMA PV Cre and 7 SMA P < P=0.0018; wt P=0.7943; wt v (Pv Chat)Cre P=0.9995; P=0.0188; sma sma v (Pv Chat)Cre P=0.0182; (Pv Chat)Cre P=0.0023; v (Pv Chat)Cre P=0.7291) P = v SMA P=0.025; wt v PvCre P=0.0180; P=0.9532) Day 1 SMA vs PV Cre P = Day 2 P = Day 3 P = Day 4 P = Day 5 P = Day 6 P = Day 7 P = Day 8 P = Day 9 P = Day 10 P = Day 11 P = F (5, 14) = F (4, 11) = Day 1 SMA vs PV Cre t = df = 8 Day 2 t =1.286 Day 3 t = Day 4 t = Day 5 t = df = 13 Day 6 t = df = 15 Day 7 t = df = 15 Day 8 t = df = 15 Day 9 t = df = 13 Day 10 t = df = 11 Day 11 t = Day 12 P = Day 12 t = df = 7 9

10 5h unpair t n = 11, 7 Righting reflex times from 11 SMA ChAT Cre, 17 SMA PVChAT Cre and 7 SMA Day 1 SMA vs ChAT Cre P = Day 2 P = Day 3 P = Day 4 P = Day 5 P= Day 6 P = Day 7 P = Day 8 P = Day 9 P = Day 10 P = Day 11 P = Day 12 P = Day 1 SMA vs PV ChAT Cre P = Day 2 P = Day 3 P = Day 4 P = Day 5 P = Day 6 P < Day 7 P < Day 8 P < Day 9 P < Day 10 P < Day 11 P < Day 1 SMA vs ChAT Cre t = df = 15 Day 2 t = Day 3 t = df = 16 Day 4 t = df = 16 Day 5 t = df = 15 Day 6 t = df = 16 Day 7 = t = df = 16 Day 8 t = df = 16 Day 9 t = df = 15 Day 10 t = df = 13 Day 11 t = Day 12 t = df = 11 Day 1 SMA vs PV ChAT Cre t = df = 22 Day 2 t = df = 22 Day 3 t = df = 22 Day 4 t = df = 22 Day 5 t = df = 21 Day 6 t = df = 22 Day 7 t = df = 22 Day 8 t = df = 21 Day 9 t = df = 19 Day 10 t = df = 16 Day 11 t = df = 15 Day 12 P < Day 12 t =

11 6a 6c 6d 6f 6g 6h 6i unpair t unpair t unpair t unpair t unpair t unpair t unpair t n = 14, 8 n = 3, 3 animals n = 3, 3 animals n = 11, 7 n = 11, 7 n = 11, 7 n = 11, 7 Righting reflex times from 14 WT, 8 PV TeNT neurons from 3 WT and 3 PV TeNT neurons from 3 WT and 3 PV TeNT PV TeNT PV TeNT PV TeNT PV TeNT Day 1 WT vs PV TeNT P = Day 2 P = Day 3 P < Day 4 P < Day 5 P = Day 6 P = Day 7 P = Day 8 P = Day 9 P = Day 10 P = Day 11 P = Day 12 P = P = WT vs PV TeNT dendritic density 050 μm P = WT vs PV TeNT dendritic density μm P = Day 1 WT vs PV TeNT t = df = 20 Day 2 t = df = 20 Day 3 t = df = 20 Day 4 t = df = 20 Day 5 t = df = 20 Day 6 t = df = 20 Day 7 t = df = 17 Day 8 t = df = 20 Day 9 t = df = 20 Day 10 t = df = 20 Day 11 t = df = 17 Day 12 t = df = 15 t = df = 27 WT vs PV TeNT dendritic density 050 μm t = df = 84 WT vs PV TeNT dendritic density μm t = df = 37 P = t = df = 16 P = t = df = 16 P = t = df = 16 P = t = df = 16 11

12 6k 7b 7d unpair t Oneway unpair t 7g pair t 7j unpair t n = 8, 6 n = 8, 6, 6, 8 n = 8, 6 n = 3, 3 n = 3, 3 PV TeNT (firing frequency), SMA, SMAPV Cre and SMAChAT Cre and PV TeNT and SMA (half width) and SMA 10 pa current WT vs PV TeNT P = pa P = pa P = pa P = pa P = pa P = pa P = pa P = pa P = P = (post hoc: wt P=0.0004; wt v PvCre P=0.2142; wt P=0.0093) 10 pa current WT vs PV TeNT t = df = 8 20 pa t = pa t = df = 9 40 pa t = pa t = pa t = pa t = df = 9 80 pa t = df = 8 90 pa t = df = 8 F (3, 25) = P = t = df = 13 P = t = df = 2 10 pa current WT vs SMA P = pa P = pa P = pa P = pa P = pa P = pa P = pa P = pa current WT vs SMA t = df = 4 20 pa t = df = 4 30 pa t = df = 4 40 pa t = df = 4 50 pa t = df = 4 60 pa t = df = 4 70 pa t = df = 4 80 pa t = df = 4 90 pa P = pa t = df = 4 12

13 8b 8c 2e 2g 2i Oneway unpair t Mann (2g) Mann (2i) unpair t n = 3 each group n = 14, 12, 8, 4 n = 4, 5 (2g): n = 4, 5 (2i): n=18,14, SMA, SMAPV Cre, SMAChAT Cre, SMA(PVChAT) Cre, Pv TeNT, SMAkainate treat Righting times from WT, SMA and Kainatetreat P4 L5 motor and SMA (2g): P4 L5 motor and SMA (2i): P2 L2 motor neurons from 3 WT and 3 SMA (2g) (2i) WT v SMA: P < ; WT v PvTeNT: P<0.001; WT v SMA ChATCre: P<0.01; SMA v SMA PvCre: SMA v SMA (Pv ChAT)Cre: SMA v SMA kainate: P< SMA vs SMAkainatetreat Day 0 P = Day 1 P = Day 2 P = Day 3 P = Day 4 P = WT vs WTkainatetreat Day 0 P = Day 1 P = Day 2 P = Day 3 P = Day 4 P = Input resistance: P = Time constant: P = Rheobase: P = (2g): P = (2i): P= DF=104 SMA vs SMAkainatetreat Day 0 t = df = 7 Day 1 t = df = 8 Day 2 t = df = 5 Day 3 t = df = 6 Day 4 t = df = 6 WT vs WTkainatetreat Day 0 t = df = 13 Day 1 t = df = 24 Day 2 t = df = 21 Day 3 t = df = 17 Day 4 t = df = 14 Input resistance: U = 9 Time constant: U = 8 Rheobase: U = 7 (2g): U = 5 (2i): t= df=30 3d Mann n = 3, 3 P3 WT, prior & after transection P = 0.20 U = 1 13

14 7d 8b Mann Oneway n = 5, 4 n = 3 each group neurons WT and SMA P11 L2 motor, SMA, SMAPV Cre, SMAChAT Cre, SMAPVChAT Cre, Pv TeNT (somatic synapses) P11 L2 motor Oneway, n = 3 SMA, SMAPV Cre, SMAChAT Cre, each SMAPVChAT 8c group Cre, Pv TeNT (dendritic synapses) Soma size: P = Input resistance: P = 0.02 Soma: P < P=0.0052; wt wt v (Pv Chat)Cre P=0.0178; sma P=0.0209; sma v (Pv Chat)Cre P=0.0586; (Pv Chat)Cre P=0.9677; v (Pv Chat)Cre P=0.0051) Dendritic density 050 μm: P < P=0.1199; wt wt v (Pv Chat)Cre P=0.6459; sma P=0.4735; sma v (Pv Chat)Cre P=0.0224; (Pv Chat)Cre P=0.9025; v (Pv Chat)Cre P=0.0013) dendritic density μm: P < P=0.0002; P=0.8352; wt wt v (Pv Soma size: U = 7 Input resistance: U = 0 Soma: F (4, 60) = Dendritic density 050 μm: F (4, 223) = dendritic density μm: F (4, 153) = dendritic density μm: F (4, 80) =

15 9e 10d 10e 11a 11e 11h 12f, 12h, 12k Mann Mann Oneway Mann Mann t Mann 12f &12h figure 12k n = 4, 4 n = 9, 14 n = 10, 11, 12 n = 3, 5 animals n = 4, 3 n = 4,3 12f & 12h: n=39 synapses /exp group 12k: n = 5, 3 P4 L5 motor and SMA Lifespan of SMA PV Cre and SMA Pv Cre Lifespan of SMA ChAT Cre and ; SMA(PVChAT) Cre and PV TeNT CMAP responses from QL P4 WT and PV TeNT P4 L5 motor and PV TeNT (12f): P5 L2 GAD65 intensity in SMA and SMABDNF (12h): P5 L2 GAD67 intensity in SMA and SMA BDNF (12k): neurons from SMA and SMAAAV9 GFPBDNF 10 pa current WT vs SMA P = pa P = pa P = pa P = pa P = pa P = pa P > pa P = pa P = > pa current WT vs SMA U = 7 20 pa U = 4 30 pa U = 5 40 pa U = 5 50 pa U = 7 60 pa U = 7 70 pa U = 8 80 pa U = 5 90 pa U = 6 P = U = 21.0 P = (post hoc: v P=0.0010; v (Pv Chat)Cre P=0.0132; v (Pv Chat)Cre P=0.4980) P = P = F (2, 30) = U = 4 U = 2 P = t=5.187 df=5 (12f): P< (12h): P= (12k): P = (12f): U = (12h): U = (12k): U = 5 15

16 12L 13b 13e 13g 14e 14f 14h unpair t Mann Pair t Mann Oneway unpair t unpair t n =8, 6,3 n = 11, 7 n = 3, 3 n = 8, 6 n = 6, 6, 9 n=3 each group n = 3 from each group Righting times from WT, SMA and SMAAAV9GFP BDNF inject and SMA and SMA and SMA P4 L5 motor, SMA and PV TeNT and SMA neurons from SMA and SMA kainatetreat 10 pa current SMA v SMA BDNF P= pA current P= pA current P= pA current P= pA current P= pA current P= pA current P= pA current P= pA current P=0.078 AHP duration: P = AHP amplitude: P 0.06 WT vs WT GxTx1E: P = SMA vs SMA GxTx1E: P = SMA vs SMA BDNFinject 10pA t = pA t = pA t = pa t = pa t = pa t = pa t = pa t = pa t = AHP duration: U = 23 AHP duration: U = 11.5 WT vs WT GxTx1E: t = df = 2 SMA vs SMA GxTx1E: t = df = 2 P = U = 2 P < P=0.9395; wt v PvTeNT p=0.0004; sma v PvTeNT p=0.0002) P= P = F (2, 18) = t=1.821 df20 t = df = 25 16

17 Representative figures 1. Are any representative images shown (including Western blots and immunohistochemistry/staining) in the paper? If so, what figure(s)? 2. For each representative image, is there a clear statement of how many times this experiment was successfully repeat and a discussion of any limitations in repeatability? If so, where is this (section, paragraph #)? Statistics and general methods 1. Is there a justification of the sample size? If so, how was it justifi? Even if no sample size calculation was perform, authors should why the sample size is adequate to measure their effect size. 2. Are statistical s justifi as appropriate for every figure? a. If there is a section summarizing the statistical methods in the methods, is the statistical for each experiment clearly defin? b. Do the data meet the assumptions of the specific statistical you chose (e.g. normality for a parametric )? Where is this describ (section, paragraph #)? c. Is there any estimate of variance within each group of data? Is the variance similar between groups that are being statistically compar? Where is this describ (section, paragraph #)? 1F; 2F; 3A; 5A; 6B; 7I; 8A,C,E; Supplementary Figs. 1A and B; 3A, B and C; 4A and B; 5AD; 6A F; 7B; 8A, 11AB; 12A, C, E, G and I; 14A, B, D, F and G. METHODS ON LINE and THIS CHECKLIST. Bas on previous experiments and publications, we have us a sample size that allow acceptable variability in order to draw valid conclusion. Yes, the appropriate statistical analysis is justifi in details in METHODS ONLINE. The specific s appli for each graph are also in the s. Yes, in Methods online section there is a paragraph entitl Statistical Analysis where we justifi each for each experiment. For each experiment the statistical is detail in the corresponding figure. Yes, as in the Statistical Analysis paragraph of Methods online, we that the D'Agostino & Pearson omnibus normality was us to assess the normality for all the data. If violat, nonparametric s were us. d. Are s specifi as one or twosid? Twosid s were us for all the experiments. e. Are there adjustments for s? Posthoc s are stat in text. No. 17

18 3. To promote transparency, Nature Neuroscience has stopp allowing bar graphs to statistics in the papers it publishes. If you have bar graphs in your paper, please make sure to switch them to dotplots (with central and dispersion statistics display) or to boxandwhisker plots to show data distributions. 4. Are criteria for excluding data points? Was this criterion establish prior to data collection? Where is this describ (section, paragraph #)? 5. Define the method of randomization us to assign subjects (or samples) to the experimental groups and to collect and process data. If no randomization was us, state so. Where does this appear (section, paragraph #)? 6. Is a statement of the extent to which investigator knew the group allocation during the experiment and in assessing outcome includ? If no blinding was done, state so. 7. For experiments in live vertebrates, is a statement of compliance with ethical guidelines/regulations includ? 8. Is the species of the animals us? 9. Is the strain of the animals (including background strains of KO/ transgenic animals us)? 10. Is the sex of the animals/subjects us? 11. Is the age of the animals/subjects? 12. For animals hous in a vivarium, is the light/dark cycle? Us align AND scatter dot plots. No blinding was done. Yes. Methods online, section: Animals (1st para). Yes Yes. Methods online, section: Animals (1st & 2nd para). Yes. Both sexes were us. (Methods online, 1st para). Yes. Throughout the manuscript and methods online. No 13. For animals hous in a vivarium, is the housing group (i.e. number of animals per cage)? No 18

19 14. For behavioral experiments, is the time of day (e.g. light or dark cycle)? 15. Is the previous history of the animals/subjects (e.g. prior drug administration, surgery, behavioral ing)? a. If behavioral s were conduct in the same group of animals, is this? 16. If any animals/subjects were exclud from analysis, is this? Reagents a. How were the criteria for exclusion defin? Where is this describ (section, paragraph #)? b. Specify reasons for any discrepancy between the number of animals at the beginning and end of the study. Where is this describ (section, paragraph #)? 1. Have antibodies been validat for use in the system under study (assay and species)? a. Is antibody catalog number given? Where does this appear (section, paragraph #)? b. Where were the validation data (citation, supplementary information, Antibodypia)? Where does this appear (section, paragraph #)? 2. Cell line identity a. Are any cell lines us in this paper list in the database of commonly misidentifi cell lines maintain by ICLAC and NCBI Biosample? No No Yes. Kv2.1 and Kv2.2 antibodies were in knockout mouse tissue. Yes, in Supplementary Table 2 b. If yes, include in the Methods section a scientific justification of their useindicate here in which section and paragraph the justification can be found. 19

20 c. For each cell line, include in the Methods section a statement that specifies: the source of the cell lines have the cell lines been authenticat? If so, by which method? have the cell lines been for mycoplasma contamination? Data availability Provide a Data availability statement in the Methods section under "Data availability", which should include, where applicable: Accession codes for deposit data Other unique identifiers (such as DOIs and hyperlinks for any other datasets) At a minimum, a statement confirming that all relevant data are available from the authors Formal citations of datasets that are assign DOIs A statement regarding data available in the manuscript as source data A statement regarding data available with restrictions See our data availability and data citations policy page for more information. Data deposition in a public repository is mandatory for: a. Protein, DNA and RNA sequences b. Macromolecular structures c. Crystallographic data for small molecules d. Microarray data Deposition is strongly recommend for many other datasets for which structur public repositories exist; more details on our data policy are available here. We encourage the provision of other source data in supplementary information or in unstructur repositories such as Figshare and Dryad. We encourage publication of Data Descriptors (see Scientific Data) to maximize data reuse. Where is the Data Availability statement provid (section, paragraph #)? Computer code/software All relevant data are available from the corresponding author. Any custom algorithm/software that is central to the methods must be suppli by the authors in a usable and readable form for readers at the time of publication. However, referees may ask for this information at any time during the review process. 1. Identify all custom software or scripts that were requir to conduct the study and where in the procures each was us. 20

21 2. If computer code was us to generate results that are central to the paper's conclusions, include a statement in the Methods section under "Code availability" to indicate whether and how the code can be access. Include version information as necessary and any restrictions on availability. Human subjects 1. Which IRB approv the protocol? Where is this stat (section, paragraph #)? 2. Is demographic information on all subjects provid? 3. Is the number of human subjects, their age and sex clearly defin? 4. Are the inclusion and exclusion criteria (if any) clearly specifi? 5. How well were the groups match? Where is this information describ (section, paragraph #)? 6. Is a statement includ confirming that inform consent was obtain from all subjects? 7. For publication of patient photos, is a statement includ confirming that consent to publish was obtain? fmri studies For papers ing functional imaging (fmri) results please ensure that these minimal ing guidelines are met and that all this information is clearly provid in the methods: 1. Were any subjects scann but then reject for the analysis after the data was collect? a. If yes, is the number reject and reasons for rejection describ? 21

22 2. Is the number of blocks, trials or experimental units per session and/ or subjects specifi? 3. Is the length of each trial and interval between trials specifi? 4. Is a block, eventrelat, or mix design being us? If applicable, please specify the block length or how the eventrelat or mix design was optimiz. 5. Is the task design clearly describ? 6. How was behavioral performance measur? 7. Is an ANOVA or factorial design being us? 8. For data acquisition, is a whole brain scan us? If, state area of acquisition. a. How was this region determin? 9. Is the field strength (in Tesla) of the MRI system stat? a. Is the pulse sequence type (gradient/spin echo, EPI/spiral) stat? b. Are the fieldofview, matrix size, slice thickness, and TE/TR/ flip angle clearly stat? 10. Are the software and specific parameters (model/functions, smoothing kernel size if applicable, etc.) us for data processing and preprocessing clearly stat? 11. Is the coordinate space for the anatomical/functional imaging data clearly defin as subject/native space or standardiz stereotaxic space, e.g., original Talairach, MNI305, ICBM152, etc? Where (section, paragraph #)? 12. If there was data normalization/standardization to a specific space template, are the type of transformation (linear vs. nonlinear) us and image types being transform clearly describ? Where (section, paragraph #)? 13. How were anatomical locations determin, e.g., via an automat labeling algorithm (AAL), standardiz coordinate database (Talairach daemon), probabilistic atlases, etc.? 22

23 14. Were any additional regressors (behavioral covariates, motion etc) us? 15. Is the contrast construction clearly defin? 16. Is a mix/random effects or fix inference us? a. If fix effects inference us, is this justifi? 17. Were repeat measures us ( measurements per subject)? a. If so, are the method to account for within subject correlation and the assumptions made about variance clearly stat? 18. If the threshold us for inference and visualization in figures varies, is this clearly stat? 19. Are statistical inferences correct for s? a. If, is this label as uncorrect? 20. Are the results bas on an ROI (region of interest) analysis? a. If so, is the rationale clearly describ? b. How were the ROI s defin (functional vs anatomical localization)? 21. Is there correction for s within each voxel? 22. For clusterwise significance, is the clusterdefining threshold and the correct significance level defin? Additional comments Additional Comments 23