Ire1 alpha

After indicated treatments, cell lysates were harvested using RIPA buffer (Cell Signaling Technologies) with protease inhibitors (cOmplete, Roche) and phosphatase inhibitors (PhosSTOP, Roche). Antibodies used were as follows: ARID1A (Santa Cruz; sc-373784), α-tubulin (Santa Cruz; sc-5286), β-Actin (C-4) (Santa Cruz; sc-47778), β-Actin (Cell Signaling; 8457), BAF57/SMARCE1 (Bethyl Laboratories, A300-810A), BAF60a (Santa Cruz; sc-135843), BAF155 (Cell Signaling; 11956), BAF170 (Santa Cruz; sc-166237), SMARCA4 (Santa Cruz; 17798), Cleaved Caspase-3 (Cell Signaling; 9664), c-MYC (Santa Cruz; sc-764) or c-MYC (Cell Signaling; 9402), cyclin B1 (Cell Signaling; 4135 and 4138), cyclin D1 (Santa Cruz; sc-718), GAPDH (Cell Signaling; 2118S and 97166S), GRP78 (Rockland Antibodies, Limerick, PA; 200–301 F36), IRE1-alpha (Cell Signaling; 3294), lamin A/C (Cell Signaling; 2032), PSMB5 (Abcam, Cambridge, MA; ab3330), SMARCB1/SNF5 (Bethyl A301-087A), UBE2C (Proteintech, Rosemont, IL; 66087–1). Results shown are representative of at least two biological replicates.

General chemicals were from VWR (West Chester, PA). Phorbol 12,13 myristate acetate (PMA) and ionomycin were from Calbiochem (Gibbstown, NJ). Antibodies to the following epitopes were sourced as follows: ATF6 (Abcam, Cambridge, MA); phospho PERK-Thr380, IRE1 alpha, Calnexin, perilipins A and B, ATG7, ATG12, LC3A, LC3B and Grb2 (Cell Signalling, Danvers, MA). Oil Red O and KLH-DNP were from EMD (Gibbstown, NJ). IgE anti-DNP were from Sigma. 4′,6-Diamidino-2-Phenylindole (DAPI), Alexa- and HRP conjugated secondary antibodies were from Invitrogen (Temecula, CA) and Amersham (Piscataway, NJ). Insulin was added at 0.01 mg/ml where indicated in the presence of 10% FBS, 0.25 micromolar Dexamethasone, 0.5 micromolar IBMX (FDI).