Fig. 4.
Inhibition of PDHC and LDH improves cell viability. (A) Western blotting for the E1α subunit of the PDHC and acetylated histone H3 (H3K9) of HeLa cells transfected with scrambled (scr) siRNA or anti-E1α siRNA (PDHA1 siRNA) before incubations with anti-human CD95-Ab (clone CH11). Histone H3 and β-actin were used as loading controls. (B) Flow cytometry histograms of cells positive (death cells, in red) or negative for propidium iodide (in black) following incubations with CD95-Ab. (C) Averages of the percentages of death cells after incubation with CD95-Ab in cells knock-down for E1α or in control cells transfected with scr siRNA (n = 5 for group). Likelihood ratio for generalized linear model: p = 2.2 × 10−16. (D) Western blotting for LDH-A and acetylated histone H3 (H3K9) of HeLa cells transfected with scr siRNA or anti-LDH-A siRNA before incubations with anti-human CD95-Ab (clone CH11). Histone H3 and β-actin were used as loading controls. (E) Flow cytometry histograms of cells positive (death cells, in red) or negative (in black) for propidium iodide following incubations with CD95-Ab. (F) Averages of the percentages of death cells after incubation with CD95-Ab in cells knock-down for LDH-A or in control cells transfected with scr siRNA (n = 5 for group). Likelihood ratio for generalized linear model: p = 8.9 × 10−10. (G) Chemical structure of galloflavin. (H) Flow cytometry histograms of cells positive (death cells, in red) or negative (in black) for propidium iodide following incubations with the CD95-Ab and galloflavin or vehicle (0.075% DMSO). (I) Averages of the percentages of death cells after incubation with CD95-Ab (clone CH11) in cells incubated with galloflavin or vehicle (n = 5 for group). Likelihood ratio for generalized linear model: p = 0.001. Ab, antibody; LDH, lactate dehydrogenase; PDHC, pyruvate dehydrogenase complex; scr siRNA, scrambled siRNA. (This figure appears in colour on the web.)