Supporting Information

Transcription

1 Supporting Information Fluorescent BODIPY-anionic boron cluster conjugates as potential agents for cell tracking Mahdi Chaari, 1,4 Nerea Gaztelumendi, 2 Justo Cabrera-González, 1# Paula Peixoto- Moledo, 2 Clara Viñas, 1 Elba Xochitiotzi-Flores, 3 Norberto Farfán, 3 Abdelhamid Ben Salah, 4 Carme Nogués, 2* Rosario Núñez 1 * 1 Instituto de Ciencia de Materiales de Barcelona (ICMAB-CSIC), Campus de la UAB, E Bellaterra (Barcelona), Spain. rosario@icmab.es 2 Departament de Biologia Cel lular, Fisiologia i Immunologia. Universitat Autònoma de Barcelona, E Bellaterra (Barcelona), Spain. carme.nogues@uab.cat 3 Facultad de Química, Departamento de Química Orgánica, Universidad Nacional Autónoma de México, México D. F., México. 4 Laboratoire des Sciences des Matériaux et de l Environnement, Faculté des Sciences de Sfax, Université de Sfax, B.P. 1171, 00 Sfax, Tunisie. M.C. and N.G. contributed equally to this work. # Current address: Department of Chemistry, Trinity College Dublin, Dublin 2, Ireland. 1

2 Table S1. Molecular weight and concentration for compounds 5-9. Table S2. Correction of the fluorescence intensity (FI) obtained by flow cytometry at 5 µg B/ml. Figure S1. Absorbance of compounds 7-9 incubated with MTT for 3 h in absence of cells (a) and formazan salts formation (b). Figure S2. GI50 of compound 5-9 Figure S3. Cellular uptake comparison between BODIPY dye and BODIPY-boron clusters at 100 µg/ml of B Figure S4. (a) 1 H NMR, (b) 11 B{ 1 H} NMR and (c) 13 C{ 1 H} NMR spectra of compound 5. Figure S5. (a) 1 H NMR, (b) 11 B{ 1 H} NMR and (c) 13 C{ 1 H} NMR spectra of compound 6 Figure S6. (a) 1 H NMR, (b) 11 B{ 1 H} NMR and (c) 13 C{ 1 H} NMR spectra of compound 7 Figure S7. (a) 1 H NMR, (b) 11 B{ 1 H} NMR and (c) 13 C{ 1 H} NMR spectra of compound 8 Figure S8. (a) 1 H NMR, (b) 11 B{ 1 H} NMR and (c) 13 C{ 1 H} NMR spectra of compound 9 2

3 Table S1. Molecular weight and concentrations for compounds 5-9. Compound MW (g/mol) Boron concentration (µg/ml) Molarity (µm)

4 Table S2. Correction of the fluorescence intensity (FI) obtained by flow cytometry at 5 µg B/ml to avoid the differences in the emission intensity due to the compound structure (FI values in arbitrary units (a.u.)). Compound FI in MEM a b Relative FI 1/Relative FI c FI (flow cytometry) d Corrected FI flow cytometry e a Intrinsec FI values obtained in cell culture medium (MEM), represented in Figure 3c. b The relative FI represents the FI of each compound with respect to compound 5 [FI(compound)/FI(5)]. c 1/relative FI represents the increment of fluorescence of compound 5 with respect to the rest of compounds. d Fluorescence intensity obtained by flow cytometry. e The corrected FI has been calculated by the product of 1/Relative FI (column 4) and FI flow cytometry (column 5). 4

5 Figure S1. Absorbance of compounds 7-9 incubated with MTT for 3 h in absence of cells (a) and formazan salt formation (b). a) Compounds 7-9 at 5 µg/ml were not able to reduce the MTT in absence of cells (control = culture medium + MTT). Absorbance values were normalized with the control medium. b) An homogenous formazan formation was observed in control cultures, whereas higher amounts of formazan salts (black precipitates) were detected in many cells incubated with compounds 7 and 9 at 5 µg/ml. Scale bar: 100 µm. 5

6 Figure S2. Fitting curve to GI50 determination of compound 5-9 and GI50 concentrations expressed in terms of boron containing or molarity 6

7 Figure S3. Cellular uptake comparison between BODIPY dye and BODIPY-boron clusters at 100 µg/ml of B. Internalization was equal or higher than BODIPY dye alone at 487 and 712 µm (molarities of the compounds at 100 µg B/ml). Mean values and SD form three independent experiments. a.u.: arbitrary units. 7

8 (a) (b)

9 (c) Figure S4 (a) 1 H NMR, (b) 11 B{ 1 H} NMR and (c) 13 C{ 1 H} NMR spectra of compound 5. 9

10 (a) (b)

11 (c) Figure S5. (a) 1 H NMR, (b) 11 B{ 1 H} NMR and (c) 13 C{ 1 H} NMR spectra of compound 6. 11

12 12 (a) (b)

13 (c) Figure S6. (a) 1 H NMR, (b) 11 B{ 1 H} NMR and (c) 13 C{ 1 H} NMR spectra of compound 7. 13

14 (a) (b)

15 5.42 (c) Figure S7. (a) 1 H NMR, (b) 11 B{ 1 H} NMR and (c) 13 C{ 1 H} NMR spectra of compound 8. 15

16 (a) (b)

17 (c) Figure S8. (a) 1 H NMR, (b) 11 B{ 1 H} NMR and (c) 13 C{ 1 H} NMR spectra of compound 9. 17