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. 2013 Jun 7;20(9):1161–1173. doi: 10.1038/cdd.2013.45

Figure 4.

Figure 4

Atg5 and Atg7 are required for GX15-070-induced autophagy and cell death. (a) TE671 and RMS13 cells were transduced with control vector (shCtrl) or vectors containing two distinct shRNA sequences against Atg5 (shAtg5#1, shAtg5#2). Expression of Atg5 was determined by western blotting. (b) TE671 and RMS13 cells were treated with 100 nM GX15-070 for the indicated time points and LC3 lipidation was assessed by western blotting. (c) TE671 and RMS13 cells were treated with the indicated concentrations of GX15-070 for 72 h. Cell viability was determined by MTT assay and is expressed as percentage of untreated controls. Data represent mean+S.D. of three independent experiments performed in triplicate; *P<0.05; **P<0.001 comparing control with Atg5 knockdown cells. (d) TE671 and RMS13 cells were transfected with control siRNA (siCtrl) or siRNA targeting Atg7 (siAtg7). Expression of RIP1 was determined by western blotting. (e) TE671 and RMS13 cells were treated with 100 nM GX15-070 for the indicated time points and LC3 lipidation was detected by western blotting. (f) TE671 and RMS13 cells were treated with the indicated concentrations of GX15-070 for 72 h. Cell viability was determined by MTT assay and is expressed as percentage of untreated controls. Data represent mean+S.D. of three independent experiments performed in triplicate; *P<0.05; **P<0.001 comparing control with Atg7 knockdown cells