Osmi 4

LNCaP, 22RV1, and C4-2 cell lines were obtained from the American Tissue Culture Collection. Cells were maintained in 10% fetal bovine serum (FBS) in RPMI medium. RNA isolation was performed using the illustraMiniSpin-kit (GE Healthcare) according to manufacturer’s instructions, and cDNA was synthesized using the qScript cDNA Synthesis Kit (Quantabio). For androgen-starvation experiments, cells were maintained in phenol red-free RPMI medium supplemented with charcoal-stripped FBS. AT7519 was purchased from Selleckchem for reverse-phase protein array and metabolite profiling experiments, and for the other experiments, from MedChemExpress. OSMI-2 was synthesized in Professor Suzanne Walker’s laboratory (Harvard Medical School), while OSMI-4 (Martin et al. 2018 (link)) and dihydrotestosterone were purchased from MedChemExpress. Cell lysates for western blotting were prepared as previously described (Itkonen and Mills 2013 (link)), and antibodies used are from Cell Signaling technology: p-Ser2-RNA Pol II (13499) and OGT (24083); from Santa Cruz Biotechnology: p-H2AX (517348); and from Abcam: RL2 (ab2739) and Actin (ab49900). Signal intensity of western blot (densitometry) was determined using Image Lab version 6.0 (Bio-Rad). Cell number was assessed using crystal violet staining assay as previously described (Barkovskaya et al. 2020 (link)).

Human MCF-7 breast cancer cells, A549 lung cancer cells, and U251 glioblastoma cells were cultured in Dulbecco's modified Eagle's medium (Sigma–Aldrich), supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. All cell lines aforementioned were purchased from the Cell Bank of Type Culture Collection of Chinese Academy of Science. EGF was purchased from R&D Systems. TEPP-46 and OSMI-4 was purchased from MedChemExpress.

Cell lines 22RV1, C4-2, LNCaP, PC3, and RWPE-1 were obtained from the American Tissue Culture Collection. PNT1 cells were acquired from Sigma. LNCaP95 (LN95) cell line was kindly provided by Professor Stephen Plymate (University of Washington). 22RV1, C4-2, LNCaP, and PNT1 cells were maintained in the RPMI medium supplemented with 10% fetal bovine serum (FBS). LN95 and RWPE-1 cells were maintained in 10% charcoal-stripped FBS in the phenol red-free RPMI media and in the keratinocyte serum-free media, respectively. Samuraciclib (Patel et al. 2018 (link)), YKL-5-124 (Olson et al. 2019 ), OSMI-4 (Martin et al. 2018 (link)), NVP2 (Olson et al. 2018 (link)), and Thiamet G (Yuzwa et al. 2008 (link)) were obtained from MedChemExpress.