Rabbit anti blimp1

Protein lysates were obtained from cells or excised tumors from transplanted mice through solubilization in lysis buffer [PBS; 1% Sodium Dodecyl Sulfate (SDS; Sigma-Aldrich 05030); PhosSTOP™ (Roche 04 906 845 001); cOmplete™ Protease Inhibitor Cocktail (Roche 11873580001)], separated on 8–15% SDS-PAGE in reducing conditions and transferred onto nitrocellulose membrane (GE Healthcare 10600002). Western blot (WB) analyses were performed following standard protocols. The following Abs were used: rat ant-IRF4 (1:500; Biolegend, 646402); rabbit anti-BLIMP1 (1:1000; Cell Signaling, 9115); rabbit anti-phospho eIF2α (1:1000; Cell Signaling, 3398); rabbit anti-eIF2α tot (1:1000; Cell Signaling, 5324); rabbit anti-phospho S6 Ribosomal Protein (1000; Cell Signaling, 5364); rabbit anti-S6 Ribosomal Protein (1000; Cell Signaling, 2317); rabbit anti-LC3B polyclonal Ab (1:500; Novus Biologicals, NB100–220); rabbit anti-phospho AKT (S473) (1:1000; Cell Signaling, 9271); rabbit anti-phospho AKT (T308) (1:1000; Cell Signaling, 2965) rabbit anti-Akt (pan) (C67E7) (1:1000; Cell Signaling, 4691); rabbit anti-phospho BAD (1:1000; Cell Signaling, 4366); rabbit anti-BAD tot (1:1000; Cell Signaling, 9239); mouse anti-ACTB/b actin (1:5000; Sigma-Aldrich, A5441). Quantifications were obtained with the gel analysis option of ImageJ software.

Lymphocytes were lysed in RIPA buffer (50 mM Tris, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 0.1% SDS, 0.5% DOC, and 1% Triton X-100), sonicated, boiled and used for SDS – PAGE followed by Western blotting. The following primary polyclonal antibodies were used: rabbit anti-IFITM3 (Proteintech, Chicago), rabbit anti-BCL-6 (Cell Signaling Technology, MA), rabbit anti-Blimp1 (Cell Signaling Technology), rabbit anti-AID (Cell Signaling Technology) and mouse anti-β-actin (Cell Signaling Technology). Goat anti-rabbit IgG-HRP and rabbit anti-mouse IgG-HRP (Santa Cruz Biotechology, Texas) were used as secondary antibodies. ECL western blotting detection reagents (Solarbio, Beijing) were used for detection.