Hnrnpa1 4b10

Protein lysates from MB primary cultures and frozen operative tissue were prepared in lysis buffer (50 mM HEPES, pH7.0, 150 mM NaCl, 2% SDS, 1 mM EDTA) supplemented with protease and phosphatase inhibitors (Roche). Proteins were separated on 4–20% Bis-Tris gradient polyacrylamide gels (Invitrogen) and transferred onto Immuno-Blot nitrocellulose membranes (Bio-Rad Laboratories). Membranes were then incubated in blocking buffer (1X TBS containing 5% milk and 0.05% Tween-20, 1 hour), probed overnight with antibodies specific for HNRNPA2/B1 (Cell Signaling, 1:1000), HNRNPA1 (4B10; Santa Cruz; 1:2000), HNRNPC1/C2 (4F4; Santa Cruz; 1:5000), cMYC (9E10; Santa Cruz; 1:500) and actin (Sigma; 1:2000).

The following antibodies were used: TRF1 (Abcam, ab371; a generous gift from Dr. Titia de Lange) for Western blots, TRF1 (affinity-purified rabbit antibody against recombinant TRF1 from serum no. 605 448) (Grolimund et al. 2013 (link)) for IF, TRF2 (Millipore, 05-521) for Western blots, γH2AX (Millipore, 05-636) for both Western blots and IF, hnRNPA1 (4B10; Santa Cruz Biotechnology, sc-32301) for Western blots, 53BP1 (Novus Biologicals, NB100-304) for IF, 53BP1 (Novus Biologicals, NB100-305) for Western blots, and phospho-ATM-Ser 1981 (Abcam, ab81292) for Western blots.