Skip to main content
. Author manuscript; available in PMC: 2018 Apr 24.
Published in final edited form as: Oncogene. 2015 Sep 14;35(18):2345–2356. doi: 10.1038/onc.2015.295

Figure 6. Androgen induces miR-135a-5p expression in PC cells.

Figure 6

A. LNCaP and VCaP cells were plated in media supplemented with 10% charcoal stripped FBS for 48 hours. Then, R1881 (1 nM) was added and the cells were incubated for an additional 24 hours. At the end of treatment, total RNA was isolated by the Trizol method and reverse transcribed. Stem loop-mediated reverse transcription of mRNA was performed with 250 ng of total RNA for miR-135a-5p and RNU6B. Relative expression of miR-135a-5p in each cell line was determined by quantitative PCR and normalized to RNU6B; Bars, S.E. B. LNCaP and VCaP cells were treated with MDV3100, as indicated, for 48 hours. Total RNA was isolated and reverse transcribed utilizing stem loop-mediated reverse transcription for miR-135a-5p and RNU6B. Relative expression of miR-135a-5p in each cell line was determined by quantitative PCR and normalized to RNU6B; Bars, S.E. C. We examined the ChIP-Seq profiles for AR, FOXA1, GATA2, SRC-2, CBP, p300, RNA Pol II, H3K4me1, H3K4me2, H3K4me3 and H3K27Ac at the RMST gene locus in LNCaP cells utilizing the IGV browser. We observed significant binding/localization of these proteins to a region approximately 5 kb downstream of the miR-135a-5p coding sequence within the RMST gene (gray arrow), which is the host gene for miR-135a-5p (black arrow).