Ar v7

For ChIP-qPCR, primers for KLK3-enh, KLK2-enh, NKX3.1-enh, SGK1-enh, SGK3-enh, MBOAT2-enh, RAB11B-FBS, PDK4-FBS, and CDK1-FBS were listed in Supplementary Table 1. Primers for TMPRSS2-enh, ZBTB16-enh, and SH2B1-enh were described previously 11 (link). Primers for TFF1-enh, NRIP1-enh, PGR-enh, and DSCAM-enh were described previously 56 (link),57 (link).
For RT-qPCR, primer and probe sets for KLK3 and ZBTB16 were listed in Supplementary Table 1. Primer and probe sets for MBOAT2 (Hs01027245_m1), ELOVL7 (Hs00405151_m1), FKBP5 (Hs01561006_m1), NKX3.1 (Hs00171834_m1), SGK1 (Hs00178612_m1), ACSL3 (Hs00244853_m1), ELOVL5 (Hs01094711_m1), NANS (Hs00219054_m1), and AR-V7 (AI6R0CI) were purchased from Thermo Fisher.

Whole cell lysate was extracted from NF-κB activated/inactivated PCa cells. A 20μg aliquot (40-60μg for AR-V7 antibody) of each protein sample was separated on a 4 to 12% Tris-glycine gradient gel (NOVEX™), and then transferred to nitrocellulose membranes (Schleicher & Schuell, Germany). The membranes were blocked with 5% skim milk in TBS-T (Trypsin buffered saline, 1% Tween-20) buffer. The AR (N20, Santa Criuz) and AR-V7 (Precision) antibodies were added (AR, 1:1000; AR-V7, 1:200) and the blots were incubated o/n in 4 C°. After washing three times for 10 minutes each in TBS-T, incubation was performed for 1 hour with the secondary horseradish-peroxidase-conjugated anti-rabbit antibody. The signals were developed by an ECL detection system (Amersham Biosciences, Amersham, USA).