Fig. 2.
TAK1 inhibition impairs mitochondria, which is restored by hydrogen sulfide but not by general ROS scavenger. (A) Tak1iKO BMDMs were treated with vehicle (ethanol, EtOH) or with 4OHT for 5 d. Some EtOH-treated cells were also treated with 300 nM 5ZOZ for 5 h before the fixation. Mitochondrial morphology was visualized with anti-TOM20 immunofluorescence staining. (B) Quantification of mitochondrial morphology changes at 5 d post EtOH or 4OHT treatment. Some EtOH-treated cells were treated with 300 nM 5ZOZ and 3 mM NAC for 5 h. (C) Different types of ROS modulators; modulators of mitochondrial electron flux, sodium sulfide (0.4 mM); cysteine (3 mM); NAC (3 mM); or a general free radical scavenger, tBHQ (20 µM), were simultaneously treated with 300 nM 5ZOZ in BMDMs for 18 h. Cells were analyzed with CM-H2DCFDA (DCFDA) and Sytox Red or with MitoSox and Sytox Green. MFIs of live cells relative to that in vehicle-treated cells (none) are shown. Effectiveness of tBHQ was verified in separate experiments shown in SI Appendix, Fig. S2. One-way ANOVA, multiple comparisons, and Tukey test; N.S., not significant (B and C).