Leibovitz s medium

All lung cell lines were obtained directly from the American Type Culture Collection (ATCC, Manassas, VA, USA). Lung SCC lines H226, H2170, H520, H596, and ChaGo‐k‐1 were maintained in RMPI‐1640 (Nacalai Tesque), Calu‐1 in McCoy's 5a (Nacalai Tesque, Kyoto, Japan), SK‐MES‐1 in DMEM (Nacalai Tesque), SW900 in Leibovitz’s medium (Gibco, Life Technologies, Carlsbad, CA, USA), H1869 in ACL‐4 (Gibco, Life Technologies), and H2066 in HITES medium (Gibco, Life Technologies). Normal lung fibroblast cells (MRC‐5, IMR‐90, and WI‐38) were cultured in EMEM (Gibco, Life Technologies). All media were supplemented with 10% fetal bovine serum, 2 mm l‐glutamine, 100 μg·mL⁻¹ streptomycin, and 100 U·mL⁻¹ penicillin. ACL‐4 and HITES were further supplemented with additional nutrients as recommended by the ATCC. All cell lines were authenticated with GenePrint^® 10 System (Promega, Fitchburg, WI, USA).
For inhibitor studies, belinostat (PXD101), bortezomib (Velcade), GDC0879, PD0325901, RDEA119, and GSK1120212 were obtained from Selleck Chemicals (Houston, TX, USA); cisplatin from Hospira (Lake Forest, IL, USA); MG‐132 from Sigma (St. Louis, MO, USA); and cetuximab was from Merck (Darmstadt, Germany).

AUT5 ((5 R)-5-ethyl-3-[6-(spiro[1-benzofuran-3,1’-cyclopropan]-4-yloxy)-3-pyridinyl]-2,4-imidazolidinedione; PubChem CID: 57410333) and AUT1 ((5 R)-5-Ethyl-3-(6-{[4-methyl-3-(methyloxy)phenyl]oxy}i-3-pyridinyl)-2,4-imidazolidinedione); PubChem CID: 53230344) were synthesized and purified by Autifony Therapeutics, LTD (Stevenage, UK)¹⁷ . Lyophilized compounds were dissolved in DMSO to make a 10 mM stock, which was kept at 4 ^oC and was stable for up to 3 months. Immediately before use, AUT5 was diluted to the desired working concentrations in ND96 containing (in mM): 96 NaCl, 2 KCl, 1.8 CaCl₂, 1 MgCl₂, 5 HEPES, 2.5 sodium pyruvate, adjusted to pH 7.4 with NaOH. Collagenase A was purchased from Sigma (St. Louis, MO), Leibovitz’s medium was purchased from Gibco (Thermo Fisher Scientific, Waltham, MA), PMA, DMSO and all other standard chemicals were purchased from Sigma-Aldrich (St. Louis, MO) (Supplementary Materials, Table S5).

The Sf9 insect cell line, a clonal isolate of IPLB-Sf21-AE cells, derived from the fall armyworm Spodoptera frugiperda [32 (link)], was maintained in TC-100 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS), penicillin G (60 µg/mL), streptomycin sulfate (200 µg/mL), and amphotericin B (0.5 µg/mL). Baby hamster kidney-derived BHK-21 cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco/Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS. Ae. albopictus-derived C6/36 cells were maintained in Leibovitz’s medium (Gibco/Thermo Fisher Scientific) containing 10% FBS. Ae. aegypti-derived Aag2 cells were maintained in Schneider’s Drosophila medium (Gibco/Thermo Fisher Scientific) containing 10% FBS. Sf9 cells, C6/36 cells, and Aag2 cells were cultured at 27 °C, and BHK-21 cells were cultured at 37 °C with 5% CO₂.