Anti hnrnp m

Protein samples were separated by sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis (PAGE) followed by gel transfer to nitrocellulose membrane (Cytiva). The membranes were sequentially incubated with primary antibodies and horseradish peroxidase (HRP)-conjugated secondary antibodies followed by detection with enhanced chemiluminescence system (Millipore) using Amersham Imager 680 (Cytiva). The primary antibodies used in this study were anti-FLAG (Sigma-Aldrich, F3165, 1:8000 diluted), anti-GAPDH (ImB, MM001, 1:3000 diluted), anti-MeCP2 (Diagenode, pAb-052-050), anti-Rbfox1(Millipore, MABE159), anti-Rbfox2 (Bethyl, A300–864A), anti-Rbfox3 (Millipore, MAB377), anti-Matrin3 (Bethyl, A300–591A), anti-hnRNP M (Santa Cruz, sc-20001), anti-hnRNP A1 (Santa Cruz, sc-32301), anti-hnRNP C (Abclonal, A0057), anti-hnRNP H1 (Abclonal, A5924), anti-hnRNP U (Abclonal, A9907), anti-hnRNP F (Abclonal, A5505), anti-NF110 (Abclonal, A2496), anti-hnRNP K (Abclonal, A1701), anti-histone H3 (Abcam, ab1791), anti-DDX5 (Abcam, ab21696), and anti-GFP (Proteintech, 66002-1-Ig). Dilutions of all primary antibodies were 1:1000 if not specifically mentioned. HRP-conjugated secondary antibodies (1:2500 diluted) were anti-mouse IgG (Promega, W4021), anti-rabbit IgG (Promega, W4011), and conformation-specific anti-rabbit IgG (Cell Signaling, 3678).

Antibodies were obtained commercially. From Santa Cruz Biotechnology: anti-AKAP95 (sc-10766, 1:500); anti-DDX5 (sc-32858, 1:500); anti-hnRNP M (sc-20002, 1:500); hnRNP F (sc-10045, 1:500); anti-His (sc-803, 1:500); and anti-RNA Pol II (sc-899, 1:500). From others: Anti-hnRNP A1 (Novus Biologicals, NB100–672, 1:2,000), anti-Pol II (Covance, MPY-127R for clones 8WG16, H14 and H5, 1:500), anti-hnRNP H (Bethyl Laboratories, A300–511A, 1:2,000); DDX17 (Bethyl Laboratories, A300–509A, 1:2,000); anti-HA (Abcam, ab9110, 1:1,000); anti-GAPDH (Chemicon, MAB374, 1:5,000); anti-FLAG (Sigma, A8592, 1:1,000); anti-FLAG (Sigma, A2220, M2 beads); and anti-MBP (New England BioLabs, E8032S, 1:5,000).