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Fig. 3 | Molecular Cancer

Fig. 3

From: A novel AR translational regulator lncRNA LBCS inhibits castration resistance of prostate cancer

Fig. 3

LBCS inhibits AR protein translation and restrains AR signaling activation. a The AR and its target genes were detected in LBCS overexpressed LNCaP-AI and LNCaP-Bic cells, LBCS knockdown LNCaP cells by real time qPCR. The results were normalized to GAPDH and presented as the means ± SD of values obtained in three independent experiments. b The protein level of AR and its target genes were detected in LBCS overexpressed LNCaP-AI and LNCaP-Bic cells, LBCS knockdown LNCaP cells by western blotting. c The concentration of PSA was validated in cultural supernant of LBCS overexpressed LNCaP-AI and LNCaP-Bic cells, LBCS knockdown LNCaP cells. The PSA concentration was adjusted by cell count of each group. The results are presented as the means ± SD of values obtained in three independent experiments. d-e LBCS was detected by ISH and AR was detected by ICH in 18 cases of PCa tissue. H-Score was calculated then analyzed by Spearman’s correlation analysis. Representative images of ISH and image were displayed. Black scale bar: 500 μm, red scale bar: 50 μm. f LBCS depletion LNCaP and control cells were treated with cycloheximide for 2, 4 and 8 h. The AR protein expression was detected by western blotting. The grey scale of the blot was analyzed by Image J. g LBCS depletion LNCaP and overexpressed LNCaP-AI cells with respective control cells were treated with either DMSO or MG132 for 8 h. The AR protein expression was detected by western blotting. The grey scale of the blot was analyzed by Image J. h Flag-AR and HA-ubiquitin vectors were co-transfected to either LBCS overexpressed or control 293 T cells. CoIP was conducted by anti-flag (AR) antibody then the product was detected using anti-HA (ubiquitin) antibody. AR expression of whole cell lysates (WCL) was detected as loading control of each group. i ChIP analysis of IgG, AR, and RNA polymerase-II status of AR target genes in LNCaP cells after LBCS knockdown. The values are normalized to input and presented as the means ± SD. (See also Additional file 13: Figure S3) *p < 0.05, **p < 0.01

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