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. 2015 Jan 21;11(2):214–224. doi: 10.4161/15548627.2014.994400

Figure 3.

Figure 3.

PARP1 promotes TNFSF10-induced autophagy by sustaining formation of the HMGB1-BECN1 complex. (A) PANC-1 cells were transfected with control shRNA and PARP1 shRNA for 48 h and then treated with TNFSF10 (100 ng/ml) for 24 h. The levels of MAP1LC3A-I/II and SQSTM1 were assayed by western blot. Relative band intensities of MAP1LC3A-II and SQSTM1 were quantified (P < 0.05). AU, arbitrary unit. (B and C) The indicated PANC-1 cells were treated with TNFSF10 (100 ng/ml) for 24 h. The number of MAP1LC3A puncta (yellow arrows) (B) was assayed by confocal microscopic analysis (* P < 0.05 vs. control shRNA group; bar = 10 μm). The interaction between HMGB1 and BECN1 (C) was assayed by coimmunoprecipitation analysis. (D) PANC-1 cells were transfected with wild-type and HMGB1C106S cDNA for 48 h and then treated with TNFSF10 (100 ng/ml) for 24 h. The levels of indicated proteins were assayed by western blot. (E) PANC-1 cells were transfected with the indicated control shRNA, PARP1 shRNA, and HMGB1C106S cDNA for 48 h and then treated with TNFSF10 (100 ng/ml) for 24 h. The levels of the indicated proteins were assayed by western blot. Relative band intensities of MAP1LC3A-II and cyt-HMGB1 were quantified. In parallel, cell viability was assayed (n = 3 , * P < 0.05). (F) PANC-1 cells were transfected with wild-type and HMGB1[3KA] cDNA for 48 h and then treated with TNFSF10 (100 ng/ml) for 24 h. The levels of indicated proteins were assayed by western blot.