Anti hnrnp k

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).

The following antibodies were used for western blot analysis in this study: anti-c-Myc (Cell Signaling Technology); anti-GAPDH and anti-β-Actin (CMC-TAG); normal rabbit IgG, normal mouse IgG_2a, anti-HUR, anti-cyclin D1, anti-CNBP and anti-NADSYN1 (Santa Cruz); anti-FLAG (Sigma-Aldrich); anti-cyclin E1, anti-CDK2, and anti-CDK4 (ImmunoWay Biotechnology Company); anti-HNRNPK (ABclonal); anti-DHCR7 (ABCAM). anti-c-Myc used for ChIP assay was from Santa Cruz. Thymidine, Nocodazole, Mimosine, EGF, hydrocortisone, Cholera Toxin, insulin and Doxycycline was from Sigma-Aldrich. Actinomycin D was from Solarbio. Strepavidin beads for RNA pull-down assay was from Invitrogen.