Ab22397

To determine the effect of PRMT5 knockdown on AR expression, PRMT5 were transiently or stably knocked down in LNCaP cells for 96 h, and total RNA was isolated using TRIzol Reagent (Thermo Fisher Scientific, Waltham, MA, USA). One microgram of total RNA was used for reverse transcription using the High Capacity cDNA Reverse Transcription Kit (Promega) according to manufacturer's instruction. The qRT-PCR analysis of AR or AR target genes (PSA, KLK2, TMPRSS2) was performed as described previously.^{30 (link)} Antibodies against AR (SC-816, Santa Cruz, CA, USA), PRMT5 (07-405, Millipore, Billerica, MA, USA), PSA (1984-1, Epitomics, Burlingame, CA, USA), FLAG (Sigma, F-1804), Sp1 (ab13370, Abcam, Cambridge, MA, USA), H4R3me2s (Abcam, ab5823), H3R8me2s (Abcam, ab130740), H2AR3me2s (Abcam, ab22397), and Brg1 (Abcam, ab110641) were used for western blotting analysis.