Fig. 1: Autophagy is required for survival of PCa cells under anoikis conditions. | Cell Death & Disease

Fig. 1: Autophagy is required for survival of PCa cells under anoikis conditions.

From: ATF4/CEMIP/PKCα promotes anoikis resistance by enhancing protective autophagy in prostate cancer cells

Fig. 1: Autophagy is required for survival of PCa cells under anoikis conditions.

A Cell survival rates in anoikis-resistant (AR) and parental (P) PC-3 and DU145 cells were shown by CCK-8 after continuous suspension and adherent culture for 7 days. B Autophagic flux in PC-3-P and PC-3-AR cells was detected by immunofluorescent staining after continuous suspension culture for 48 h, and endogenous LC3B tagged with a tandem fluorescent-mCherry-GFP as a reporter. Autophagy flux were detected in 2D culture, and a total of more than 20 cells under each condition were counted for quantification of autophagy (Scale bar, 50 μm). C Transmission electron microscopy manifested that PC-3-AR cells showed more double-membrane autophagosomes compared with PC-3-P cells (Original magnification, ×1000, ×1600, respectively). D Western blot analysis of the expression level of autophagy-related proteins (Beclin1, p-Beclin1, LC3BII/LC3BI) in PCa-AR (PC-3-AR、DU145-AR) and PCa-P (PC-3-P、DU145-P) cells. E Western blot analysis of the expression level of autophagy-related proteins (Beclin1, p-Beclin1, LC3BII/LC3BI) in PC-3 cells at different suspension time points. F Western blot analysis of the expression level of autophagy-related proteins (Beclin1, p-Beclin1, LC3BII/LC3BI) in PC-3-P and PC-3-AR cells after the treatment of Rapa (200 nM) or 3-MA (10 μM). G Flow cytometry assay certified the detachment-induced apoptosis rate in parental and anoikis-resistant PC-3 and DU145 cells after the treatment of Rapa (200 nM) or 3-MA (10 μM) in suspension situation for 48 h. (Data are presented as the means ± SD of three independent experiments; *p < 0.05, **p < 0.01, ***p < 0.001).

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