Infliximab

Five human NSCLC cell lines with EGFR activating mutations (PC-9 [ECACC #90071810], H1975 [ATCC #CRL-5908], II-18 [#RCB2093; RIKEN BioResource Research Center, Tsukuba, Japan], HCC827 [ATCC #CRL-2868], and HCC4006 [ATCC #CRL-2871]) and one human NSCLC cell line WT for EGFR (A549 [ATCC #CCL-185]) were studied. The EGFR and TP53 mutation status of each cell line is provided in Supplementary Table 1. PC-9, H1975, II-18, HCC827, and HCC4006 cells were cultured in RPMI 1640 medium (Gibco, Carlsbad, CA, USA), and A549 cells were maintained in DMEM (Gibco). Each medium was supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin (Gibco). All cells were maintained under a humidified atmosphere of 5% CO₂ and at 37 °C. Osimertinib (#S7297; Selleck Chemicals, Houston, TX, USA) was dissolved in DMSO (Fujifilm Wako Pure Chemical Industries, Osaka, Japan) and stored at −20 °C. Infliximab (Mitsubishi Tanabe Pharma, Osaka, Japan) and doxycycline (#631311; Takara Bio, Shiga, Japan) were stored at −20 °C. JSH-23 (#S7351, Selleck Chemicals) was dissolved in DMSO (Fujifilm Wako Pure Chemical Industries) and stored at −20 °C. Recombinant human TNF-α (#210-TA-005; R&D Systems, Minneapolis, MI, USA) was dissolved in sterile phosphate-buffered saline (PBS) containing 0.1% bovine serum albumin and was stored at −20 °C.

Infliximab (IFX), Etanercept (ETN), Certolizumab Pegol (CZP) were provided by Mitsubishi Tanabe Pharma Corporation (Osaka, Japan), Takeda Pharmaceutical Company Limited (Osaka, Japan) and UCB Japan (Tokyo, Japan), respectively. Methotrexate and folic acid were obtained from Wako Chemicals (Osaka, Japan). Rho kinase inhibitor, Y27632, and PKH-26 were supplied from Sigma (St. Louis, MO, United States). TO-PRO-3 was from Thermo Fisher Scientific (Waltham, MA, United States). Rabbit anti-phospho JNK monoclonal antibody (mAb) and anti-JNK polyclonal antibody (Ab) were purchased from cell signaling technology (Danvers, MA, United States). Anti-GAPDH Ab was from Gene Tex (Irvine, CA, United States).