For whole-cell lysis, harvested cell culture pellets were rinsed with phosphate-buffered saline (PBS), lysed in SDS-containing buffer [4% SDS, 20% glycerol, 0.1 M tris-HCl (pH 7.5)], and sonicated. Protein lysates were centrifuged at maximum speed for 10 min. Protein extracts were quantified using the BCA Protein Assay Kit (Pierce, 23225), and equal amounts of protein were resolved by SDS–polyacrylamide gel electrophoresis (PAGE) electrophoresis and blotted onto the transfer membrane (Immobilon-P, Merck Millipore, IPVH00010). Membrane blocking (10% bovine serum albumin/tris-buffered saline and 0.2% Tween 20 for 30 min at room temperature) was followed by incubation with the appropriate primary antibodies and horseradish peroxidase–conjugated secondary antibodies (Cell Signaling Technology). Proteins were detected by enhanced chemiluminescence (Thermo Fisher Scientific). The following primary antibodies were used: Anti-monomethylated arginine (MeR4-100, Cell Signaling Technology, #8015, 1:1000 dilution), anti-symmetric dimethylated arginine (Cell Signaling Technology, #13222, 1:1000 dilution), and anti-asymmetric dimethylated arginine (Cell Signaling Technology, #13522, 1:1000 dilution) were purchased from Cell Signaling Technology; anti-vinculin (06-866, 1:10,000 dilution) was purchased from Merck Millipore.

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