Fig. 3: LINC00675 stabilizes androgen receptor (AR) protein by block its ubiquitination.

a AR expression levels in different prostate cancer (PCa) cell lines. b AR protein expression was downregulated in LINC00675-depleted LNCaP-SF and LNCaP-C4-2b cells. c AR mRNA expression was not changed in LINC00675-depleted LNCaP-SF and LNCaP-C4-2b cells. Data are represented as mean ± SD. n = 3. d LNCaP-SF and LNCaP-C4-2b cells were transfected with siRNA to knock down LINC00675. After 24 h, the cells were treated with 20 μM MG132 for 6–8 h and harvested. Western blot showed that AR expression was rescued after treatment. e LINC00675-depleted LNCaP-SF and LNCaP-C4-2b cells were treated with 100 mg/ml CHX and harvested at different time points as indicated. AR’s half-life is shortened after LINC00675 depletion. CHX cycloheximide. f, g LINC00675 blocks ubiquitination of AR confirmed by exogenous and endogenous Co-IP assay, detected by western blot. f 293T cells were transfected with Flag-AR, His-Ub, pLINC00675 plasmids, or siLINC00675 as indicated. g LNCaP-SF and LNCaP cells were transfected with siLINC00675 or pLINC00675 and relative controls, and then treated with MG132 (20 μM) for 6–8 h before harvest. TCL total cell lysate.