Figure 1 | Cell Death & Disease

Figure 1

From: Downregulation of ASPP2 improves hepatocellular carcinoma cells survival via promoting BECN1-dependent autophagy initiation

Figure 1

ASPP2 silencing is important for induction of autophagy in HCC cells. (a) HepG2 and HCC-LM3 liver cancer cells were incubate in CM (0 h) or EBSS for 4, 8, 12, and 24 h. Cells were collected for western blotting with antibodies. (b) HCC-LM3 cells were incubated in CM or in EBSS for 8 h. Cells were stained for ASPP2 or LC3 and imaged by immunofluorescence microscopy. Scale bars: 5 μm. (c and e) HepG2 and HCC-LM3 were infected with LV-shNon or LV-shASPP2 for 72 h; Huh7 and HCC-LM3 were transfected with pcDNA3.1 or pASPP2 for 48 h. (c) Transmission electron microscopy showed formation of autophagosomes after EBSS treatment for 6 h in HCC cells. The scale bars represent 1 μm. (d) Representative images of GFP-LC3 puncta (autophagosomes) in HCC cells cultured in EBSS for 6 h with or without 3-MA (10 mM, 1 h) pretreatment. Quantitation of autophagy (with GFP-LC3 punctate dots) in conditions shown in lower panel. The scale bars represent 10 μm. (e) Western blots analysis of HCC cells cultured in EBSS for 6 h with or without 3-MA (10 mM, 1 h) pretreatment. (f) HepG2 infected with LV-shNon or LV-shASPP2 were treated with EBSS for indicated time with or without CQ. Band intensity was quantified using ImageJ. Data represented the mean±S.D. from triplicate experiments. (*P<0.05; **P<0.01). CM, completed medium; EBSS, serum-free Earle’s Balanced Salt Solution medium; 3-MA, 3-methyladenine; CQ, chloroquine

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