Fig. 4

Knockdown of ANRIL affects cells proliferation and apoptosis by mediating miR-191. The expression vectors of sh-ANRIL#1, sh-ANRIL#2 and miR-191 mimic were transfected into HepG2 cells to alter ANRIL and miR-191 expression. a The relative expression level of miR-191 in miR-191 mimic transfected cells was determined by qRT-PCR assay. b Cell viability was examined in HepG2 cells after transfection with sh-ANRIL#1, sh-ANRIL#2 and miR-191 mimic by using CCK-8 assay. c and d Protein levels of CyclinD1, p53 and p21 in these transfected cells were detected by western blot assay. e Cell apoptosis and f the protein levels of pro-Caspase-3/− 9 and cleaved-Caspase-3/− 9 were respectively determined by flow cytometry and western blot. ANRIL: CDKN2B antisense RNA 1; miR-191: microRNA-191; qRT-PCR: quantitative real-time reverse-transcription polymerase chain reaction; CCK-8: Cell Counting Kit-8; *P < 0.05; **P < 0.01, ***P < 0.001