Beta actin a1978

DMEM (Dulbecco’s Modified Eagle’s Medium) was obtained from Invitrogen (Carlsbad, CA). Fetal bovine serum (FBS) was obtained from Hyclone Laboratories (Logan, UT). Cell-culture plates were purchased from Denville Scientific (Charlotte, NC). Celecoxib and Orlistat were obtained from Cayman Chemical (Ann Arbor, MI). Cell Titer 96 Aqueous One Solution Cell Proliferation assay was obtained from Promega (Madison, WI). The enhanced chemiluminescence reagents (ECL) kit and Pierce BCA protein assay were obtained from ThermoScientific (Rockford, IL). BD Pharmigen FITC Annexin V Apoptosis Detection Kit I was obtained from BD Biosciences (San Jose, CA). Control, MUC2, and CHOP shRNA lentiviral particles were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). For western blotting Beta-Actin (A1978) was obtained from Sigma Aldrich, St. Louis, MO; PARP-1 (CST 46D11), GRP78 (CST C50B12), ATF4 (CST D4B8), PUMA (CST D30C10), NOXA (CST D8L7U), CHOP (CSTL63F7), and Cleaved Caspase 3(CST Asp175) were obtained from Cell Signaling Technologies (Danvers, MA). SYTOX Orange for nucleic acid labeling was obtained from Life Technologies (Grand Island, NY). Anti-rabbit and anti-mouse horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Jackson Immunology Research (West Grove, PA).

Antibodies to IKKβ (8943), IKKα (2682), p-p53 S15 (9284), p-p53 S392 (9281), p-p65 (3033), p65 (3987), p-Iкbα (2859), and Iкbα (4814) were from Cell Signaling (Danvers, MA, USA), beta actin (A1978) was from Sigma (St. Louis, MO, USA), and p53 DO-1 (SC126) was from Santa Cruz Biotechnology (Dallas, TX, USA). Recombinant active GST- IKKβ (31176) was purchased from Active Motif (Carlsbad, CA, USA). Recombinant GST-p53 (14-865) was purchased from Millipore (Temecula, CA, USA). Recombinant GST-IкBα (P323-30G) was purchased from SignalChem (Richmond, BC, Canada). pcDNA3.0-p53wt (69003), pCMV2-Flag- IKKβ (11103) were purchased from Addgene (Cambridge, MA, USA). pUNO1-HA-IKKβ was purchased from InvivoGen (puno1ha-hikkb). pcDNA3.0-Flag-p53 S392A was custom ordered from SignalChem. Doxorubicin (Doxo) (D2141) was purchased from Sigma (St. Louis, MO, USA). TPCA-1 (15115) was purchased from PerkinElmer (Waltham, MA, USA). Lipofectamine RNAiMAX and Lipofectamine 2000 transfection reagents were purchased from Life Technologies (Carlsbad, CA, USA). TRC human IKKβ shRNA (TRCN0000018917) was purchased from Dharmacon (Lafayetter, CO, USA) and pLKO.1 scramble (1864) was purchased from Addgene.