Figure 2

Downregulation of ASPP2 promotes starvation-induced autophagy through regulating BECN1. (a) HepG2 and HCC-LM3 cells 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. After cultured in EBSS, cells were performed for the analysis BECN1 mRNA and protein level with quantitative PCR (left) and western blot (right). (c) Huh7 cells were co-transfected with pEGFP-LC3 and pBECN1 or pASPP2 into for 48 h, then were cultured in EBSS for 6 h. Representative images of GFP-LC3 (upper) and quantitation of the autophagic cells (lower). The scale bars represent 10 μm. (d) Huh7 cells transfected and treated as (c), but without pEGFP-LC3 were lysed and subjected to western blotting. (d) HCC-LM3 cells infected with LV-shNon or LV-shASPP2 for 72 h were transfected with siBECN1 for 48 h, and then were incubated in EBSS for 6 h. Cells lysates were subjected to western blotting. Data represented the mean±S.D. from triplicate experiments. (*P<0.05). 3-MA, 3-methyladenine