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. 2023 May 4;14(5):304. doi: 10.1038/s41419-023-05836-7

Fig. 1. Upregulation of CHRM4 is associated with prolonged androgen withdrawal.

Fig. 1

A CHRM4 and AR protein levels of LNCaP, 22Rv1, C4-2, C4-2-MDVR, PC3, and LASCPC01 cells, measured by a western blot analysis. B, C CHRM4 mRNA abundances in LNCaP and C4-2 cells during 1, 2, 3, 4, and 5 months of 20 μM MDV3100 treatment, measured by an RT-qPCR analysis. * vs. parental LNCaP or C4-2 cells, by a one-way ANOVA. D Relative CHRM4 mRNA levels of C4-2 cells cultured in charcoal-stripped serum (CSS)-containing medium for 5 and 10 days, followed by treatment with 10 nM dihydrotestosterone (DHT) for 24 h. Quantification of relative mRNA levels is presented as the mean ± SEM of three biological replicates. *p < 0.05, **p < 0.01, ***p < 0.0001. E CHRM4, androgen-responsive markers (KLK3 and NKX3-1), and AR protein levels in C4-2 cells cultured in CSS-containing medium for 5 and 10 days, followed by treatment with 10 nM DHT for 24 h. F CHRM4, KLK3, NKX3-1, and AR protein levels of LNCaP and C4-2 cells cultured in 20 μM MDV3100 for 1 week. G Relative mean expressions of the AR, KLK3, NKX3-1, and CHRM4 in LNCaP cells from 3 weeks to 11 months of androgen withdrawal (ADT) in the GDS3358 database. * vs. the control, by a one-way ANOVA. H GSEAs of TCGA prostate dataset revealing negative associations between high CHRM4 expression in prostate tissues with gene signatures representing androgen-responsive signaling (GO, Nelson, Wang, PID, and Hallmark). NES normalized enrichment score, FDR false discovery rate.