Anti-rabbit primary antibodies for Lipin-1 (Abcam, ab181389) and Lipin-2 were purchased from Abcam (Cambridge, UK). The anti-rabbit primary antibody for Lipin-3 was purchased from Biorbyt (Cambridge, UK). The anti-mouse primary antibody for GAPDH was purchased from Sigma-Aldrich (St. Louis, MO). The anti-mouse and anti-rabbit secondary antibodies were purchased from Promega (Madison, WI) (Supplemental Table 2). HepG2 cells were purchased from Sigma-Aldrich (St. Louis, MO) and used the same standard cell culture media and care as the prostate cell lines. Whole-cell lysates were prepared using RIPA buffer (50 mM Tris-HCl [pH 7.4], 1 M NaCl, 1% NP-40, 0.5% sodium deoxycholate, 0.1% SDS, and ddH2O) and 1% protease inhibitor cocktail from Cell Signaling Technology (Danvers, MA). Lysates were then vortexed and frozen at − 80 °C. Samples were run in triplicate (n = 3). Proteins (5 μg) were resolved by 10% SDS-PAGE and transferred onto nitrocellulose membranes with a pore size of 0.45 μm (Thermo Scientific). Membranes were blocked with 5% milk powder (AppliChem) in 1X TBST for 1 h at room temperature. Incubations with primary antibodies at 4 °C overnight were followed by incubations with the appropriate secondary antibodies at room temperature for 1 h and detection by Pierce™ SuperSignal™ West Pico PLUS Chemiluminescent Substrate from Thermo Scientific (Waltham, MA). Images were captured using the FluorChem® HD2 Imager from Alpha Innotech (San Leandro, CA). Densitometry to quantify protein expression was performed using the software FluorChem® HD2 software from Alpha Innotech (San Leandro, CA). Lipin expression was normalized to GAPDH expression.

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