P erk1 2

The cell culture media 1640 and McCoy S 5A were purchased from Gibco (Gibco, USA). The rabbit or mouse monoclonal anti-human antibodies used were ARNO (Abcam, ab56510), pEGFR (Py1068, Epitomics, 1138-1), pERK1/2 (T202/Y204, Bioworld, BS5016), EGFR (Cell Signaling, 3197), GAPDH (Bioworld, AP0063), IGF-IR (Abcam, ab39675), pIGF-IR (Abcam, ab39398), pIRS (Abcam, ab52167), pAKT (Abcam, ab106693), pIRS1 (Abcam, ab66153), pShc (Abcam, ab155170), and Ki-67(Cell Signaling, 9027). Other reagents and equipment used were as follows: SecinH3 (Merck-565725/sc-203260), siRNA oligo (Genephama), MTT (sigma, m5655), DMSO (sigma, D5879), human EGF (Peprotech, AF-100-15), human IGF-1 (Peprotech, AF-100-11), FBS (Gibco, USA), and 0.25% trypsin (Sigma), immunohistochemical kit(Zhongshan,China).

The following reagents were used: rHMGB1 (Cat No. #1690-HMB-050, R&D Systems, Minneapolis, MN, USA), G1 (Cat. No. 3577/10, TOCRIS, Bioscience, Bristol, UK), G15 (Cat. No. 3678/10, TOCRIS, Bioscience, Bristol, UK). E2, TAM, were obtained from Sigma–Aldrich (St. Louis, MO, USA). U0126, and the LY294002 were purchased from Millipore (Temecula, CA, USA). E2 was dissolved in ethanol and other drugs were solubilized in dimethyl sulfoxide (DMSO; Sigma–Aldrich). The following antibodies were used: anti-HMGB1 neutralizing antibody (Cat No. H00003146-M08, R&D systems, Minneapolis, MN, USA); p-ERK1/2 (Cat No. AP0484P, diluted 1:1000), ERK1/2 (Cat No. BS90472, diluted 1:1000), Akt (Cat No. BS6473, diluted 1:1000) were purchased from Bioworld (St Louis Park, MN, USA). p-Akt (Cat No. 4060, Cell signaling Technology, USA, diluted 1:1000), LC3B (Cat No. 2775 Cell Signaling Technology, diluted 1:1000), P62 (Cat No. 5114, Cell Signaling Technology, diluted 1:1000); Beclin1(Cat No. 210498, diluted 1:1000), GPR30 (Cat No. ab39142, diluted 1:250), HMGB1 (Cat No. ab18256, diluted 1:500), Bcl-2 (Cat No.32124, diluted 1:1000), BAX (Cat No. 182734, diluted 1:1000) were purchased from Abcam (Cambridge, MA, USA), and β-actin (Cat No. TA-09, diluted 1:1000) from Zhongshan Golden Bridge (Beijing, China).

GE (Figure 7A) and SIN (Figure 7B) (purity > 98.0%) were provided by the National Institute for the Control of Pharmaceutical and Biological Products (Beijing, China). The enzyme-linked immunosorbent assay (ELISA) kits for IFN-γ, IL-17, IL-4 and TGF-β1 were supplied from Elabscience Biotechnology Co., Ltd. (Wuhan, China). Rabbit mAb JNK, p-JNK, ERK1/2, p-ERK1/2, p38, p-p38, and β-actin were obtained from Bioworld Technology Inc. (Nanjing, China). HRP-conjugated goat anti-rabbit antibodies were purchased from ZSGB-BIO (Beijing, China). JNK inhibitor (SP600125), ERK inhibitor (PD98059), and p38 inhibitor (SB203580) were obtained from Calbiochem-Novabio chem Corp. (San Diego, CA, USA). Lipopolysaccharide (LPS), Cell counting kit-8 (CCK8) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were offered from Sigma Chemical Co. (St. Louis, MO, USA); Dulbecco’s modified Eagle’s medium (DMEM) was purchased from Thermo Scientific Co. (Waltham, MA, USA). All other chemicals used in this study were of analytical grade and were obtained from commercial sources.

Afatinib, adriamycin and cisplatinum were purchased from Meilun Biology Technology Co., Ltd (Dalian, China). Paclitaxel was obtained from Tianfeng Technology Co., Ltd (Xi'an, China). Rhodamine 123, MTT, verapamil, lipopolysaccharide (LPS), PDTC, LY294002 and other chemicals were purchased from Sigma Chemical Co. (St. Louis, MQ, USA).
The monoclonal antibody against ABCB1 was obtained from Santa Cruz Biotechnology Inc. (CA, USA). Antibodies against iκBα, p-iκBα, p65 and Lamin B1 were purchased from Epitomics, Inc. (California, USA). Antibodies against EGFR, p-EGFR, HER-2, p-HER-2, AKT, p-AKT, p38, p-p38, ERK1/2, p-ERK1/2, JNK, p-JNK and GAPDH were obtained from Bioworld Technology, Inc. (Minnesota, USA). The HRP-conjugated goat anti-rabbit secondary antibody and the CY3-conjugated goat anti-rabbit secondary antibody were purchased from Abcam Inc. (Cambridge, MA, USA).