Chop sc 575

For immunoblotting, we obtained S6K (sc-230), PERK (sc-13073) and p-PERK (sc-32577) antibodies from Santa Cruz Biotechnology, Dallas, TX, human Sestrin2 (10795-1-AP) antibody from Proteintech, Chicago, IL, BiP (3177), CHOP (2895), p-Thr389 S6K (9234), p-Ser235/236 S6 (2211), S6 (2317), p-Thr37/46 4E-BP (2855), 4E-BP (9452), p-Ser473 AKT (9273) and AKT (4691) antibodies from Cell Signaling Technology, Danverse, MA, actin (JLA20) antibody from Developmental Studies Hybridoma Bank (DSHB, Iowa city, IA), and tubulin (T5168) antibody from Sigma, St. Louis, MO. Mouse Sestrin2 antibody was described (Ro et al, 2014b (link)). For immunostaining, we obtained p53 (sc-6243), PCNA (sc-7907), β-catenin (sc-59737), CHOP (sc-575) from Santa Cruz Biotechnology, F4/80 (mf48000) from Invitrogen, Carlsbad, CA, BiP (3177), γ-H2AX (2577), p-Ser235/236 S6 (2211) and p-Thr37/46 4E-BP (2855) from Cell Signaling Technology. Azoxymethane (AOM), dextran sulfate sodium (DSS), rapamycin, 5-fluorouracil (5-FU) and irinotecan (CPT-11) were from Sigma.

Sections were permeabilized with 0.1% triton-X, blocked with normal goat serum, and antigen retrieval was performed using citric acid buffer. Primary antibodies were applied at 1:40 (CHOP; sc-575; Santa Cruz). Appropriate fluorescently-tagged secondary antibodies (647 nm) were applied, followed by DAPI staining.