Supplementary Figure 1: To test the role of mir-17~92 in orthologous genetic model of ADPKD, we generated Ksp/Cre;Pkd1 F/F (Pkd1-KO) and Ksp/Cre;Pkd1
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- Kimberly Hardy
- 5 years ago
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1 Supplementary Figure 1: To test the role of mir-17~92 in orthologous genetic model of ADPKD, we generated Ksp/Cre;Pkd1 F/F (Pkd1-KO) and Ksp/Cre;Pkd1 F/F ;mir-17~92 F/F (Pkd1-miR-17~92KO) mice. (A) Q-PCR analysis revealed that compared to control kidneys (black), Pkd1 expression was equally reduced in both Pkd1-KO (red) and Pkd1-miR-17~92KO (green) kidneys, indicating that a similar level of Cre/loxP recombination was observed. (B) Q-PCR analysis revealed that, compared to control kidneys, mir-17 expression was increased in Pkd1-KO kidneys, whereas its expression was reduced in Pkd1-miR- 17~92KO mice. (C) Kidney-weight-to-body-weight (KW/BW) ratio and the expression of kidney injury markers (D) Kim1 and (E) Ngal was reduced in Pkd1-miR-17~92KO compared to Pkd1-KO kidneys. (F) The number of cyst epithelia undergoing apoptosis did not change in Pkd1-miR-17~92KO compared to Pkd1-KO kidneys. Error bars indicate SEM. * indicates P<0.05, ns indicates P>0.05. One-way ANOVA, Tukey s multiple comparisons test (A, B, D, E), Student s unpaired t-test (C, F) 1
2 Supplementary Figure 2: To test the role of mir-17~92 in an orthologous genetic model of ADPKD bearing mutations in Pkd2, we generated Pkhd1/Cre;Pkd2 F/F (Pkd2-KO) and Pkhd1/Cre;Pkd2;miR- 17~92 F/F (Pkd2-miR-17~92KO) mice. (A) Q-PCR analysis revealed that, compared to control kidneys (black), Pkd2 expression was equally reduced in both Pkd2-KO (orange) and Pkd2-miR-17~92KO (blue) kidneys, indicating that a similar level of Cre/loxP recombination was observed. (B) Western Blot analysis and quantification of Polycystin-2 (PC-2) expression in Pkd2-KO and Pkd2-miR-17~92KO kidneys is shown. mir-17 deletion did not affect PC-2 expression in Pkd2-miR-17~92KO kidneys. (C) Q- PCR analysis revealed that, compared to control kidneys, mir-17 expression was increased in Pkd2-KO kidneys, whereas its expression was reduced by ~60% in Pkd2-miR-17~92KO mice. (D) Cyst index and the expression of kidney injury markers (E) Kim1 and (F) Ngal was reduced in Pkd2-miR-17~92KO compared to Pkd2-KO kidneys. (G) The number of cyst epithelia undergoing apoptosis was higher in Pkd2-miR-17~92KO compared to Pkd2-KO kidneys. However, this difference was not statistically significant. Error bars indicate SEM. * indicates P<0.05, ns indicates P> One-way ANOVA, Tukey s multiple comparisons test (A, C, E, F), Student s unpaired t-test (B, D, G) 2
3 Supplementary Figure 3: To test the role of mir-17~92 in a long-lived ADPKD model, we generated Ksp/Cre;Pkd1 Flox/RC (Pkd1 F/RC SKO) and Ksp/Cre;Pkd1 Flox/RC ;mir-17~92 F/F (Pkd1 F/RC DKO) mice. This compound Pkd1-mutant mouse harbors a germline hypomorphic mutation on one allele and a Cre/loxP recombination-induced, renal tubule-specific, somatic null mutation on the other allele. (A) Q-PCR analysis revealed that Pkd1 expression was not different between control (black), Pkd1 F/RC SKO (red) and Pkd1 F/RC DKO (green) kidneys, suggesting that deleting one floxed allele does not affect the total Pkd1 mrna expression. (B) In contrast, Q-PCR analysis revealed that, compared to control kidneys, mir-17 expression was increased in Pkd1 F/RC SKO kidneys, whereas its expression was reduced by ~80% in Pkd1 F/RC DKO kidneys. (C) H&E staining of kidney sections from three-week-old and five-month-old Pkd1 F/RC SKO and Pkd1 F/RC DKO mice is shown. (D&E) Kidney-weight-to-body-weight (KW/BW) ratio, (F) blood urea nitrogen (BUN) level and the expression of (G) Kim1 and (H) Ngal was reduced in Pkd1 F/RC DKO compared to Pkd1 F/RC SKO mice. Thus, deletion of mir-17~92 markedly attenuates disease progression in Ksp/Cre;Pkd1 Flox/RC mice. Error bars indicate SEM. * indicates P<0.05, ns indicates P>0.05. One-way ANOVA, Tukey s multiple comparisons test (A, B, G, H), Student s unpaired t-test (D, E, F) 3
4 Supplementary Figure 4: To test the role of mir-17~92 in a slow cyst growth ADPKD model, we generated and characterized 6-month-old Pkd1 RC/RC ; Ksp/Cre (Pkd1 RC/RC ) and Pkd1 RC/RC ; Ksp/Cre;miR- 17~92 F/F mice (Pkd1 RC/RC DKO). Unlike the preceding three ADPKD models, Cre/loxP recombination is not required to induce Pkd1 mutation, and these mice develop significant renal fibrosis. Ksp/Cre was introduced into the cross to delete mir-17~92. (A) Q-PCR analysis revealed that, compared to control kidneys, mir-17 expression was increased in Pkd1 RC/RC kidneys, whereas its expression was reduced in Pkd1 RC/RC DKO compared to Pkd1 RC/RC mice. (B) Kidney-weight-to-body-weight ratio, (C) BUN and the expression of (D) Kim1 and (E) Ngal was decreased in Pkd1 RC/RC DKO compared to Pkd1 RC/RC mice. (F) Kidney sections were stained with an antibody against Fsp1 (fibroblast-specific protein 1). Fsp1 expression (green) was decreased in Pkd1 RC/RC DKO kidneys compared to Pkd1 RC/RC kidneys. (G-J) Q-PCR analysis showed that the expression of other fibrosis markers (Col4a6, Col1a1, Acta2, and Vimentin) was also decreased in Pkd1 RC/RC DKO kidneys compared to Pkd1 RC/RC kidneys. Collectively, these results indicate that mir-17~92 deletion attenuates disease progression in Pkd1 RC/RC mice. Error bars indicate SEM. * indicates P<0.05, ns indicates P>0.05. One-way ANOVA, Tukey s multiple comparisons test (A) Student s unpaired t-test (B-J). Scale Bar: 50 µm. 4
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6 Supplementary Figure 5: (A) Genomic organization of the mir-17~92 and its paralogous clusters mir- 106a~363 and mir-106b~25 is shown. Collectively, these three mirna clusters encode mirnas that belong to either the mir-17 (blue), mir-19 (green), mir-18 (purple) or mir-25 (orange) mirna families. We have previously shown that our anti-mir-17 compound inhibits mirnas that are marked with an X in cultured cells. (B) The ability of this compound to inhibit mir-17 in vivo was measured using the mirna polysome shift assay (mipsa) 1. Normal mice were treated with a single subcutaneous dose of vehicle (PBS), anti-mir-17 (0.3, 3 or 30 mg per kg dose) or control oligonucleotide (30 mg per kg dose), and kidneys were harvested 7-days later (n=5-7 each group and dose). mir-17 fraction (normalized to let-7d) in high molecular weight (HMW) was measured by Q-PCR. mir-17 was displaced from HMW polysomes in kidneys of mice injected with anti-mir-17 in a dose-dependent manner. Similar mir-17 displacement was not observed in mice treated with vehicle or a control oligonucleotide. (C) Normal adult mice were injected with 300 mg per kg dose of anti-mir-17, vehicle or a control oligonucleotide known to produce liver toxicity (positive control). Measurement of the liver (ALT and AST) and kidney function (BUN) showed that anti-mir-17 does not produce acute liver or kidney toxicity. (D-F) The design of the delivery and therapeutic efficacy trials are shown. Error bars indicate SEM. * indicates P<0.05, ns indicates P>0.05. One-way ANOVA, Tukey s multiple comparisons test (B) Student s unpaired t-test (C) 6
7 Supplementary Figure 6: Primary cyst epithelial cultures derived from kidneys of ADPKD patients were transfected with anti-mir-17 (dose: 3 nm, 10 nm or 30 nm) or three different control oligonucleotides (dose: 30 nm, control oligo1, 2 and 3) or mock-transfected. These cells were then cultured to measure proliferation and in-vitro cyst formation. (A) Data for proliferation (n=5 for each treatment and dose) using primary cultures derived from ADPKD Donor1, 2, 3 and 5. (B) In-vitro cyst formation (n=3 for each treatment and dose) using primary cultures derived from ADPKD Donor 3. Anti-miR-17 reduced cell viability and cyst count in a dose-dependent manner. Error bars indicate SEM. *indicates P<0.05, **indicates ns indicates P<0.01, ***indicates ns indicates P<0.005, and ****indicates P< One-way ANOVA, Tukey s multiple comparisons test 7
8 Supplementary Figure 7: RNA-Seq analysis was performed to compare mrna expression profiles between kidneys of 21-day-old control, Pkd1 F/RC SKO and Pkd1 F/RC DKO mice (n=3). The differentially expressed genes (P<0.05, FDR<0.1) were further analyzed using the Ingenuity Pathway Analysis (IPA) software. Several mitochondrial pathways were differentially expressed. In particular, the oxidative phosphorylation pathway (shown above) was downregulated in Pkd1 F/RC SKO compared to control kidneys. In contrast, this pathway was upregulated in Pkd1 F/RC DKO compared to Pkd1 F/RC SKO kidneys. Green indicates downregulated genes, whereas red indicates upregulated genes. 8
9 Supplementary Figure 8: RNA-Seq analysis was performed to compare mrna expression profiles between kidneys of 21-day-old control, Pkd2-KO and Pkd2-miR-17~92KO mice (n=5). The differentially expressed genes (P<0.05, FDR<0.1) were further analyzed using the Ingenuity Pathway Analysis (IPA) software. Similar to the Pkd1-mutant mice, several mitochondrial pathways were differentially expressed. The oxidative phosphorylation pathway (shown above) was downregulated in Pkd2-KO compared to control kidneys. In contrast, this pathway was upregulated in Pkd2-miR-17~92KO compared to Pkd2-KO kidneys. Green indicates downregulated genes, whereas red indicates upregulated genes. 9
10 Supplementary Figure 9: RNA-Seq analysis was performed to compare mrna expression profiles between kidneys of 21-day-old control, Pkd1F/RCSKO and Pkd1F/RCDKO mice (n=3). The differentially expressed genes (P<0.05, FDR<0.1) were further analyzed using the Ingenuity Pathway Analysis (IPA) software to identify the upstream regulators that may be responsible for the gene expression changes observed after mir-17~92 deletion. PPARA-regulated gene network was predicted to be robustly activated after mir-17~92 deletion. Accordingly, a large number of direct PPARA targets (shown above) were upregulated in Pkd1F/RCDKO compared to Pkd1F/RCSKO kidneys. Green indicates downregulated genes, whereas red indicates upregulated genes. 10
11 Supplementary Figure 10: RNA-Seq analysis was performed to compare mrna expression profiles between kidneys of 21-day-old control, Pkd2-KO and Pkd2-miR-17~92KO mice (n=5). The differentially expressed genes (P<0.05, FDR<0.1) were further analyzed using the Ingenuity Pathway Analysis (IPA) software to identify the upstream regulators that may be responsible for the gene expression changes observed after mir-17~92 deletion in Pkd2-KO mice. Similar to Pkd1-mutant kidneys, PPARA-regulated gene network was predicted to be robustly activated after mir-17~92 deletion in Pkd2-KO kidneys. A number of direct PPARA targets (shown above) were upregulated in Pkd2-miR-17~92KO compared to Pkd2-KO kidneys. Green indicates downregulated genes, whereas red indicates upregulated genes. 11
12 Supplementary Figure 11: To test whether reduced Ppara gene dosage is sufficient to enhance proliferation and promote cyst formation, 6-week old Ppara -/-, Pkd1 RC/RC, and Pkd1 RC/RC ; Ppara-KO mice were characterized. Q-PCR analysis (A) Kim1 and (B) Ngal in the indicated mouse models is shown. Error bars indicate SEM. Multiplicity adjusted P values using Tukey s multiple comparison test are shown. 12
13 Supplementary Figure 12: Characterization of cultured cells transfected with (A) mir-17 mimics and (B) PPARA-expressing plasmid (pppara) is shown. To determine whether mir-17 affects OXPHOS, Seahorse XF 24 Analyzer was used to measure real-time mitochondrial oxygen consumption rate (OCR) of renal epithelial cells. (C) Real-time OCR tracings, (D) Basal OCR, (E) Oligomycin-treated OCR, and (F) ATP-dependent OCR of indicated cell lines is shown. ATP-dependent OCR was reduced in mimcd3 cells treated with mir-17 mimics compared to scramble mimics. Reexpression of Ppara prevented mir- 17-induced reductions in ATP-dependent OCR. (G&H) MTT assay was used to determine whether increased Ppara gene dosage inhibits mir-17-stimulated proliferation of renal epithelia. Treatment with mir-17 mimic increased proliferation of mimcd3 cells. (G) Transfection with a Ppara-expressing plasmid (pppara) reduced mir-17-stimulated proliferation whereas transfection with a control plasmid (pcmx) had no effect (n=3 biological replicates with three technical replicates). (H) Treatment with WY14643, a PPARA agonist, also reduced mir-17-stimulated proliferation whereas treatment with DMSO had no effect (n=3 biological replicates with three technical replicates). mir-17 mimic also increased proliferation of Pkd2 -/- kidney epithelia. Similar to mimcd3 cells, WY14643 reduced mir-17-stimulated proliferation of Pkd2 -/- cells whereas treatment with DMSO had no effect (n=3 biological replicates with three technical replicates). One-way ANOVA, Tukey s multiple comparisons test (D-H) Student s unpaired t-test (A, B) 13
14 Supplementary Figure 13: Song et al 2, have previously performed cdna microarrays (Human Genome U133 Plus 2.0 Array, Affymetrix) using RNA from three normal renal cortical samples, five minimally cystic ADPKD tissue samples and 13 ADPKD cysts of varying sizes (small, medium, and large). Analysis of this publically-available data ( revealed that PPARA (red arrow heads) and its targets are downregulated in human ADPKD cysts. 14
15 Supplementary Figure 14: The full, original Western blots are shown accompanied with indicated protein name (top left) and molecular weight (top right). 15
16 Supplementary Table 1:Differential expression of mirnas in ADPKD models Pkd1-KO vs. Control Pkd2-KO vs. Control mirna P10 AvgExp Fold-change P-value mmu-mir-29a E E mmu-mir-126-3p E E mmu-mir-200c E E mmu-mir E E mmu-mir E E mmu-let-7g E E mmu-mir-20a/b E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-196b E E mmu-mir E E mmu-let-7i E E mmu-mir-199a-5p E E mmu-mir-29b E E mmu-mir-30e E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-126-5p E E mmu-mir-26b E E mmu-mir E E mmu-mir-199a-3p E E mmu-mir-30c E E mmu-mir-106a+mmu-mir E E mmu-mir E E mmu-mir-362-5p E E mmu-mir E E mmu-mir-340-5p E E mmu-mir-151-5p E E mmu-mir E E mmu-mir E E mmu-mir-200a E E mmu-mir-148b E E mmu-mir E E mmu-mir E E mmu-mir-345-5p E E mmu-mir p E E mmu-mir-196a E E mmu-mir-99a E E mmu-mir E E mmu-mir-532-5p E E mmu-mir E E mmu-mir E E mmu-mir-200b E E FDR P21 AvgExp Fold-change P-value FDR 16
17 mmu-mir E E mmu-mir-188-5p E E mmu-mir-135a E E mmu-mir E E mmu-mir E E mmu-mir-301a E E mmu-mir-10b E E mmu-mir-434-3p E E mmu-mir E E mmu-mir-376b E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-26a E E mmu-mir E E mmu-mir-376c E E mmu-mir-23a E E mmu-mir E E mmu-mir-335-5p E E mmu-mir-181c E E mmu-mir E E mmu-mir-23b E E mmu-let-7d E E mmu-mir E E mmu-mir-101a E E mmu-mir E E mmu-let-7a E E mmu-mir-615-3p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-142-3p E E mmu-mir-125a-3p E E mmu-mir-148a E E mmu-mir-551b E E mmu-mir E E mmu-mir E E mmu-mir-15a E E mmu-mir E E mmu-mir-130b E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-338-3p E E mmu-mir E E mmu-let-7f E E mmu-mir-335-3p E E mmu-mir E E mmu-mir E E mmu-mir-139-5p E E mmu-mir-324-5p E E mmu-mir E E mmu-mir-450a-5p E E mmu-mir-331-3p E E mmu-mir-202-5p E E
18 mmu-mir-30b E E mmu-mir E E mmu-mir-434-5p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir p E E mmu-mir E E mmu-mir-770-3p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-669e E E mmu-mir-18a E E mmu-mir E E mmu-mir-290-5p E E mmu-mir E E mmu-mir E E mmu-let-7b E E mmu-mir-30a E E mmu-mir-146a E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-743a E E mmu-mir-190b E E mmu-mir-342-3p E E mmu-mir-34a E E mmu-mir-466c-5p E E mmu-mir-574-3p E E mmu-mir-409-5p E E mmu-mir E E mmu-mir-574-5p E E mmu-mir-181a E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-501-5p E E mmu-mir E E mmu-mir-369-3p E E mmu-mir E E mmu-mir E E mmu-mir-450b-3p E E mmu-mir E E mmu-mir E E mmu-mir-296-5p E E
19 mmu-mir E E mmu-mir-133a E E mmu-mir E E mmu-mir E E mmu-mir-331-5p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-19a E E mmu-mir-30d E E mmu-mir-380-3p E E mmu-mir E E mmu-mir-466d-3p E E mmu-mir-208b E E mmu-mir-129-5p E E mmu-mir E E mmu-mir-501-3p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-466d-5p E E mmu-mir-487b E E mmu-mir E E mmu-mir E E mmu-mir-216a E E mmu-mir E E mmu-mir E E mmu-mir-876-5p E E mmu-mir E E mmu-mir E E mmu-mir-337-5p E E mmu-mir-342-5p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-669h-5p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-466l E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-743b-3p E E mmu-mir E E mmu-mir E E mmu-mir-301b E E mmu-mir-449a E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-666-3p E E
20 mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-290-3p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-19b E E mmu-mir-130a E E mmu-mir E E mmu-mir-466a/b-3p E E mmu-mir E E mmu-mir-323-5p E E mmu-mir E E mmu-mir-106b E E mmu-mir-423-3p E E mmu-mir-532-3p E E mmu-mir-7b E E mmu-mir-582-5p E E mmu-mir-423-5p E E mmu-mir E E mmu-mir-133b E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-7a E E mmu-mir-202-3p E E mmu-mir-297c E E mmu-mir E E mmu-mir E E mmu-mir-181b/d E E mmu-mir-542-5p E E mmu-mir-875-5p E E mmu-mir E E mmu-mir E E mmu-mir-101b E E mmu-mir-193b E E mmu-mir E E mmu-mir-876-3p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-125b-3p E E mmu-mir E E mmu-mir-29c E E
21 mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-92b E E mmu-mir E E mmu-mir-669a E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-467f E E mmu-mir E E mmu-mir-669o E E mmu-mir E E mmu-mir E E mmu-mir-340-3p E E mmu-mir-362-3p E E mmu-mir E E mmu-mir E E mmu-mir p E E mmu-mir E E mmu-mir-27b E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-34b-3p E E mmu-mir E E mmu-mir-466g E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-542-3p E E mmu-mir E E mmu-mir-467c E E mmu-mir E E mmu-mir E E mmu-mir-466k E E mmu-mir p E E mmu-mir-125a-5p E E mmu-mir-467e E E mmu-mir-292-5p E E mmu-mir-669j E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-151-3p E E mmu-mir-99b E E mmu-mir E E mmu-mir-467h+mmu-mir-669d/l E E mmu-mir E E mmu-mir-669f E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-671-3p E E
22 mmu-mir E E mmu-mir-467g E E mmu-mir-323-3p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-509-5p E E mmu-mir-467a E E mmu-mir p E E mmu-mir-146b E E mmu-mir E E mmu-mir-376a E E mmu-mir E E mmu-mir-129-3p E E mmu-mir-883b-3p E E mmu-mir-338-5p E E mmu-mir-770-5p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-let-7c E E mmu-mir E E mmu-mir-142-5p E E mmu-mir-15b E E mmu-mir-125b-5p E E mmu-mir E E mmu-mir E E mmu-mir-27a E E mmu-mir-875-3p E E mmu-mir-369-5p E E mmu-mir-675-5p E E mmu-mir E E mmu-mir E E mmu-mir-1946a E E mmu-mir E E mmu-mir-34b-5p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-883b-5p E E mmu-mir E E mmu-mir E E mmu-mir-883a-3p E E mmu-mir E E mmu-mir-764-3p E E mmu-mir-764-5p E E mmu-mir-669i E E mmu-mir E E mmu-mir-3470a/b E E mmu-mir E E mmu-mir E E mmu-let-7e E E mmu-mir E E mmu-mir E E mmu-mir E E
23 mmu-mir-654-3p E E mmu-mir E E mmu-mir-337-3p E E mmu-mir-467b E E mmu-mir E E mmu-mir-302d E E mmu-mir-743b-5p E E mmu-mir E E mmu-mir-292-3p E E mmu-mir E E mmu-mir E E mmu-mir-465b-5p E E mmu-mir E E mmu-mir-92a E E mmu-mir-450b-5p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-135b E E mmu-mir-139-3p E E mmu-mir E E mmu-mir E E mmu-mir-188-3p E E mmu-mir p E E mmu-mir E E mmu-mir p E E mmu-mir E E mmu-mir E E mmu-mir-18b E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-1946b E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-208a E E mmu-mir E E mmu-mir E E
24 mmu-mir-216b E E mmu-mir E E mmu-mir E E mmu-mir-291a-3p E E mmu-mir-291a-5p E E mmu-mir-291b-3p E E mmu-mir-291b-5p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-296-3p E E mmu-mir-297b-3p E E mmu-mir-297b-5p E E mmu-mir E E mmu-mir-302a E E mmu-mir-302b E E mmu-mir-302c E E mmu-mir E E mmu-mir E E mmu-mir-324-3p E E mmu-mir E E mmu-mir E E mmu-mir-339-3p E E mmu-mir-339-5p E E mmu-mir E E mmu-mir E E mmu-mir-345-3p E E mmu-mir-34c E E mmu-mir E E mmu-mir-380-5p E E mmu-mir E E mmu-mir-384-3p E E mmu-mir-384-5p E E mmu-mir-409-3p E E mmu-mir E E mmu-mir-449b E E mmu-mir-449c E E mmu-mir-450a-3p E E mmu-mir E E mmu-mir E E mmu-mir-465a-3p E E mmu-mir-465a-5p E E mmu-mir-465c-5p E E mmu-mir-466a/e-5p E E mmu-mir-466f-5p E E mmu-mir-466h E E mmu-mir-466i E E mmu-mir-466j E E mmu-mir-467d E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-509-3p E E
25 mmu-mir E E mmu-mir-540-3p E E mmu-mir-540-5p E E mmu-mir E E mmu-mir E E mmu-mir-582-3p E E mmu-mir-590-3p E E mmu-mir-590-5p E E mmu-mir E E mmu-mir E E mmu-mir-615-5p E E mmu-mir E E mmu-mir-654-5p E E mmu-mir-666-5p E E mmu-mir-669g E E mmu-mir-669m E E mmu-mir-671-5p E E mmu-mir-673-3p E E mmu-mir-673-5p E E mmu-mir-675-3p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-693-3p E E mmu-mir-693-5p E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir E E mmu-mir-878-3p E E mmu-mir-878-5p E E mmu-mir E E mmu-mir-883a-5p E E
26 Supplementary Table 2:Commonly Dysregulated mirnas in ADPKD models. UP mmu-mir-182 mmu-mir-183 mmu-mir-96 mmu-mir-21 mmu-mir-223 mmu-mir-199a-3p mmu-mir-199a-5p mmu-mir-200c mmu-mir-100 mmu-mir-350 mmu-let-7i mmu-mir-20a+mmu-mir-20b mmu-mir-28 mmu-mir-106a+mmu-mir-17 mmu-mir-532-5p mmu-mir-143 DOWN mmu-let-7g mmu-mir-99a mmu-mir-126-3p mmu-mir-151-5p mmu-mir-196a mmu-mir-26b mmu-mir-126-5p mmu-mir-30c mmu-mir-185 mmu-mir-30e mmu-mir-194 mmu-mir-378 mmu-mir
27 Supplementary Table 3: CT values of polysome shift assay (PSA) High Molecular Weight Fraction 3 High Molecular Weight Fraction 4 Kidney PSA Score mir-17 Let-7d mir-17 Let-7d Treatment Ct Mean Ct Mean dct ddct Ct Mean Ct Mean dct ddct Average ddct Fraction 3+4 PBS PBS PBS PBS PBS PBS PBS PBS anti-mir-17 (0.3mg/kg) anti-mir-17 (0.3mg/kg) anti-mir-17 (0.3mg/kg) anti-mir-17 (0.3mg/kg) anti-mir-17 (0.3mg/kg) anti-mir-17 (3mg/kg) anti-mir-17 (3mg/kg) anti-mir-17 (3mg/kg) anti-mir-17 (3mg/kg) anti-mir-17 (3mg/kg) anti-mir-17 (30mg/kg) anti-mir-17 (30mg/kg) anti-mir-17 (30mg/kg) anti-mir-17 (30mg/kg) anti-mir-17 (30mg/kg) Control (30mg/kg) Control (30mg/kg) Control (30mg/kg) Control (30mg/kg) Control (30mg/kg) Control (30mg/kg) Control (30mg/kg) Control (30mg/kg)
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