Helenalin

Dulbecco’s modified Eagle’s medium (DMEM)/F-12 and fetal bovine serum were purchased from Thermo Fisher Scientific (Grand Island, NY, USA). BioTrace^TM NT membrane was purchased from Pall Life Sciences (Ann Arbor, MI, USA). The enhanced chemiluminescence (ECL) Western blotting detection system was purchased from Perkin Elmer (Waltham, MA, USA). Anti-phospho-EGFR (Tyr¹¹⁷³, Cat#4407), anti-phospho-p38 MAPK (Thr¹⁸⁰/Tyr¹⁸², Cat#9211), anti-phospho-JNK1/2 (Thr¹⁸³/Tyr¹⁸⁵, Cat#9255), anti-phospho-FoxO1 (Ser²⁵⁶, Cat#9461), and anti-phospho-p65 (Ser⁵³⁶, Cat#3033) antibodies were purchased from Cell Signaling (Danvers, MA, USA). Antibodies of anti-TNFR1 (Cat#sc-52739), anti-TNFR2 (Cat#sc-8041), and p38 MAPK inhibitor (p38i) VIII were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-GAPDH (Cat#MCA-1D4) was purchased from EnCor Biotechnology (Gainesville, FL, USA). AG1478, SP600125, and Tanshinone IIA were purchased from Enzo Life Science (Farmingdale, NY, USA). AS1842856 was obtained from EMD Millipore (Billerica, MA, USA). Helenalin was purchased from Cayman Chemical (Ann Arbor, MI, USA). Recombinant human TNF-α protein was purchased from R&D Systems (Minneapolis, MN, USA). TRIzol reagent, enzymes, and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA).

FBP1 wildtype (wt) and FBP1 mutant (G260R) constructs were kindly provided by Dr. Haojie Huang from the Mayo Clinic [11 (link)]. The pCMV4a-Flag-c-Myc plasmid was purchased from Addgene. FBP1 nes mutants were generated by adding a nuclear export sequence (LALKLAGLDIGS) to the FBP1 c-terminal using the KOD-Plus- Mutagenesis Kit (Toyobo, Japan). Bacterial expression vectors for GST-TRIM28 recombinant proteins were generated using the pGEX-4 T-1 backbone vector. FBP1 antibody (ab109732)was purchased from Abcam (working dilution 1:1000); beta-Tubulin (2128S) was from Cell Signaling Technology - (working dilution 1:5000); TRIM28 (ab10483) was from Abcam (working dilution 1:3000); c-Myc (5605P) was from Santa Cruz Biotechnology (working dilution 1:1000); BRD2 (ab139690) was from Abcam (working dilution 1:1000); BRD3 (A302-368A) was from Bethyl Laboratories (working dilution 1:1000); BRD4 (ab128874) was from Abcam (working dilution 1:1000). JQ1, MG132, I CBP112, puromycin and cycloheximide (CHX), were purchased from Sigma-Aldrich (Shanghai, China); MK 2206, Trametinib, GSK126, Ku55933, SB203580 and Palbociclibwere from Selleckchem (Houston, USA). Gemcitabine was obtained from Eli Lilly and Company (Indianapolis, USA). Helenalin was purchased from Cayman Chemical (Ann Arbor, USA).