Figure 7

R248Q mediates inhibition of macroautophagy induced by DCA. (a) p53 knockdown partially attenuates the inhibitory effect of DCA on the autophagy pathway. NB4 cells were transfected with control siRNA or siRNA targeting p53 and forty hours later were incubated with 10 mM DCA for a further 24 h. The effect on autophagy gene expression was analyzed by RT-qPCR. (b) Autophagy flux is restored after R248Q down-regulation. NB4 cells were transfected with control siRNA or p53 siRNA. 72 h later they were incubated with 10 mM DCA for 24 h. Autophagy flux was monitored by detection of LC3-II under CQ treatment. Graphs represent the quantification of the western blots of 3 experiments. (c) The presence of R248Q after DCA treatment causes a reduction of autophagy flux. HL60 cells were transfected with plasmids expressing wt p53 or mt p53. After 24 h transfection, cells were incubated with 10 mM DCA for 24 h. Autophagy flux was monitored by the addition of 50 μM CQ for 4 h before collecting the cells for western blot analysis. Graphs represent the quantification of the western blots of 3 experiments.