Fig. 7

HULC inhibits PTEN through ubiquitin–proteasome system mediated by autophagy-P62. a Western blotting with anti-PTEN and RT-PCR with PTEN primer in Hep3B cell line. β-actin as internal control. b Western blotting with anti-PTEN and RT-PCR with PTEN primer in Hep3B cell lines transfected with pCMV6-A-GFP plus pCMV6-XL5-PTEN (FL), pCMV6-A-GFP-HULC plus pCMV6-XL5-PTEN (FL),respectively. c (left) Co-Immunoprecipitation (IP) with anti-62 followed by Western blotting with anti-PTEN in Hep3B cell lines transfected with pCMV6 -A-GFP plus pCMV6 -XL5-PTEN (FL), pCMV6-A-GFP-HULC plus pCMV6 -XL5-PTEN (FL), pCMV6-A-GFP-HULC plus pGFP-V-RS--Sirt1 plus pCMV6-XL5-PTEN (FL), and pCMV6-A-GFP-HULC plus pLV-miR15a plus pCMV6-XL5-PTEN (FL), respectively. (right) density analysis of band. d (left) Co-Immunoprecipitation (IP) with anti-HA followed by Western blotting with anti-PTEN in the Hep3B cell line. INPUT refers to Western blotting with. Anti-PTEN. (right) density analysis of band. e (left) Co-Immunoprecipitation (IP) with anti-HA followed by Western blotting with anti-PTEN in the Hep3B cell line. (right) density analysis of band. f (left) Cells were incubated with 50 μM MG132 (Sigma) for 6 h at proper after transfection. Western blotting with anti-PTEN in Hep3B cell lines transfected with pCMV6-A-GFP plus pCMV6-XL5-PTEN (FL). g. The assay of PTEN 3'-UTR luciferase activity in Hep3B cells infected with pCMV6-A-GFP or pCMV6-A-GFP-HULC