Poly 1 c

Human HT-29, HCT116, SW480, SW620 and LOVO CRC cell lines were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). HT-29 and LOVO cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco, Grand Island, NY, USA), and the HCT116, SW480 and SW620 cells were cultured in Dulbecco's modified Eagle medium (DMEM, Hyclone, Logan, UT) supplemented with 10% fetal bovine serum (FBS) (Biological Industries, Beit Haemel, Israel) and 1% antibiotics penicillin/streptomycin (Invitrogen, Waltham, MA). The cells were cultured at 37 °C in a 5% CO₂ humidified atmosphere. Rapamycin (RAPA, Apexbio, Houston, TX), 5-fluorouracil (5-FU, Sigma Aldrich, St. Louis, MO) and amlexanox (Abcam, Cambridge, England) were dissolved in dimethyl sulfoxide (DMSO, Sigma Aldrich). Poly (I:C) (Apexbio) was dissolved in H₂O.

Ibrutinib was purchased from Macklin (Shanghai, China). Poly I:C was purchased from APExBIO (Houston, TX, USA). LPS was purchased from Sigma (St Louis, MO, USA). Mouse IL-1β, IL-6, and TNF-α ELISA kits were procured from Solaibao (Beijing, China). Anti-myeloperoxidase mouse mAb, anti-IL1 beta rabbit pAb, anti-TNF-α rabbit pAb, anti-CD8 mouse mAb, and anti-Ly6g rabbit pAb were obtained from Servicebio (Wuhan, China). APC Anti-Mouse Ly6G Antibody [1A8] and PerCP/Cyanine5.5 Anti-Mouse CD45 Antibody [30-F11] were purchased from Elabsciense (Shanghai, China). Antibodies against BTK/p-BTK (Tyr223), ERK/p-ERK, AKT/p-AKT, FLT3/p-FLT3, and EGFR/p-EGFR were bought from Cell Signaling Technology (Beverly, MA, USA). Antibodies against GAPDH, NFκB/p-NFκB, IL-1β, IL-6, IFN-γ, and TNF-α were purchased from Affinity Biosciences (Inc., Cincinnati, OH, USA). The RIPA lysis buffer and a Bradford protein assay kit were purchased from Biyuntian Biotechnology (Shanghai, China). PBS was purchased from Zhongshan Jinqiao Biotechnology (Co., Ltd., Beijing, China). A SuperLumia ECL HRP kit was procured from Abbkine (Redlands, CA, USA).

The human LUAD cell line A549 was maintained in RMPI 1640 medium (Hyclone, USA) supplemented with 10% fetal bovine serum (FBS, Hyclone, USA), 100 units/ml of penicillin, and 100 μg/ml of streptomycin at 37 °C, in a 5% CO₂ humidified incubator. Cells were harvested after treatment for 24 h with 100 μg/ml of poly(I:C) (Apexbio, USA).