Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2021 Dec 15;12:7308. doi: 10.1038/s41467-021-27077-y

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 4 — a Western blot analysis of AR or AR/MYC overexpression in LNCaP cells. LN, LNCaP; LA, LNCaP-AR; LAM, LNCaP-AR-MYC. The experiment was repeated independently three times with similar results. LNCaP-AR (LA, for AR overexpression) and LNCaP-AR-MYC (LAM, for AR and MYC overexpression) were cultured in androgen-depleted medium and treated with DHT for indicated time-points. Total RNA was subjected to real-time RT-PCR analyses. MYC-5’-UTR was based on primers targeting MYC-5’ UTR region; MYC-E2-E3 was based on primers spanning MYC exon-2 and exon-3 regions (b). The relative expression of MYC and AR targets was also determined by RT-qPCR (c–e). f Gene Set Enrichment Analysis (GSEA) using upregulated genes in siCtrl vs. siMYC (in complete FBS medium), DHT-activated genes, and androgen-repressed genes as gene sets (all from VCaP cells) to determine whether these two gene sets were regulated by MYC overexpression in LNCaP-AR cells. g RT-qPCR to determine MYC, KLK3, and TMPRSS2 mRNA levels in LA and LAM cells in full FBS medium. h ChIP-qPCR to determine AR and H3K27ac signal at KLK3 and TMPRSS2 enhancers in LA and LAM cells within full FBS medium. i Western blot analysis of MYC knockdown in VCaP cells. The experiment was repeated independently three times with similar results. j RT-qPCR to determine MYC, KLK3, and TMPRSS2 mRNA levels in VCaP cells in full FBS medium. k ChIP-qPCR to determine AR and H3K27ac signal at KLK3 and TMPRSS2 enhancers in VCaP cells in full FBS medium. For b–e, g, h, j, and k, data are presented as mean values ± SD; for each test, n = 3 biologically independent samples. For b–e, P values were determined by Wilcoxon Rank Sum Test and the multiple tests were further corrected by Bonferroni-Holm method; N.S., not significant; *, adjusted P < 0.05; **, adjusted P < 0.01; ***, adjusted P < 0.001. For g, h, j, and k, P values were calculated by two-sided Student’s t test; N.S., not significant; *, P < 0.05; **, P < 0.01; ***, P < 0.001. Source data are provided in Histogram and Immunoblot Source Data files.