Article Figures & Data
Figures
- Fig. 1.
(A) Western blot showing knockdown of SULT4A1 in primary neurons. (B) CellRox fluorescence in primary cortical neurons transduced with control shRNA or SULT4A1 shRNA. Scale bar, 20 μm. (C) Quantification of CellRox fluorescence intensity. Data represent mean ± S.E.M., n = 5, ***P < 0.001 vs. control (Student’s t test). (D) Western blots showing SULT4A1 expression in SH-SY5Y cells with or without transduction with control virus or SULT4A1 lentivirus. (E) Subcellular fractionation in SH-SY5Y showing mitochondrial localization of SULT4A1. TOM20 is used as a mitochondrial marker and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is cytosolic marker. (F) Immunostaining pictures of SULT4A1 with orthogonal projections in SH-SY5Y show that SULT4A1 (green) localize with mitochondrial marker TOM20 (red). Colocalization was determined by Pearson’s correlation coefficient, R = 0.66.
- Fig. 2.
Mitochondrial function in SH-SY5Y cells expressing SULT4A1. (A) Representative data showing mitochondrial OCR in SH-SY5Y cells transduced with control or SULT4A1 virus in presence or absence of H2O2. Quantification of basal respiration (B), maximal respiration (C), ATP turnover (D), spare respiration capacity (E), and proton leak (F). Data are mean ± S.E.M., n = 18–22 for each group. Experiments were repeated three times with similar results. (G) Confocal images showing TMRE intensity in SH-SY5Y cells transduced with control and SULT4A1 virus in presence or absence of H2O2, Scale bar, 20 μm. (H) Time-lapse quantification of TMRE fluorescence intensity before and after CCCP addition. (I) Bar graph showing quantification of TMRE intensity change (∆TMRE) before and after CCCP addition in SH-SY5Y cells. Data represents mean ± S.E.M., n = 3. (J) Cell viability in SH-SY5Y cells in presence or absence of H2O2. Data are mean ± S.E.M., n = 8. *P < 0.05; **P < 0.01; ***P < 0.001 vs. control, was calculated using one-way ANOVA and Tukey’s post hoc test.
