Plasmid extraction kit

Molecular manipulation was performed according to a standard protocol for Gram-negative bacteria. E. coli APEC40 (WT) genomic DNA and plasmid DNA were extracted using a genome extraction kit (Sangon Biotech, Shanghai, China) and plasmid extraction kit (Transgen, Beijing, China), respectively. PCR amplification was conducted by using Taq or PrimeSTAR^® Max DNA Polymerase (Takara Bio Inc., Dalian, China). A gel purification kit was used to purify the PCR products and DNA restriction fragments (Transgen, Beijing, China). DNA restriction endonuclease (Thermo Fisher Scientific, Waltham, MA, USA) digestion and homologous recombination (Vazyme, Nanjing, China) were conducted using standard methods. The primer sequences used are listed in Table 2.

DMEM medium, fetal bovine serum, T4 ligase, T7 endonuclease, Escherichia coli DH5α, plasmid extraction kit, genomic DNA extraction kit, reverse transcription kit, and SYBR^® Green RT-PCR Master Mix were purchased from TransGen Biotech Co (Beijing). ACCUTASE™ cell digestion solution was purchased from eBioScience Inc. Western blot antibodies, STAT1 (14994), p-STAT1 (Tyr701, 7649P). STAT3 (9132), p-STAT3 (Tyr705, 9145), STAT5 (9363p), p-STAT5 (Y694, 4322). AURKA (14475), p-AURKA (Thr288, 3079), PD-L1 antibody (13684), p-AKT (Ser473, 4060), AKT (pan) (4691), p-ERK/ERK antibody (4370), GAPDH (5174), β-tubulin (2128) were purchased from Cell Signaling Technology, CD3 histochemical antibody (100219), CD8 histochemical antibody (104705) were purchased from Biolegend, and 4% paraformaldehyde were purchased from Wuhan Seville Biological LTD. DAB histochemical staining kit was purchased from Zhongshan Jinqiao Biological Technology LTD (Beijing). ECL luminescence color reagent was purchased from Healthcare. Trizol RNA extraction reagent was purchased from Sigma-Aldrich (T9424). Anti-mouse PD-L1 Antibody was purchased from BioXcell.