Fig. 1

Two rounds of screening to identify the genes involved in regulating inflammasome activation in vitro. a. J774A.1 cells were pre-treated with LPS (1 μg/mL, 5 h) and then infected with C50336 TnpSC189 transposon mutants at an MOI of 20 for 4 h. The ratio of cell death was evaluated by the release of LDH in supernatants of infected cells. The Z score was calculated for each individual well in a 48-well cell plate, and a Z score ≤ − 2 or ≥ 2 was considered significant. Twenty-nine mutants induced significantly higher cytotoxicity levels (red, Z score ≥ 2) and 77 mutants induced significantly lower cytotoxicity levels (blue, Z score ≤ − 2). b. The transposon insertion sites of each candidate transposon mutants. Horizontal arrows indicate the direction of gene expression. The green genes represent the candidate genes, the blue genes represent the upstream genes of the candidate genes, the yellow genes represent the downstream genes of the candidate genes. The numbers represent the initial or terminal position in the Salmonella Enteritidis genome of each gene. The red vertical arrows represent the transposon insertion site of each candidate transposon mutants. c. J774A.1 cells were pre-treated with LPS (1 μg/mL, 5 h) and then infected with candidate transposon mutants at an MOI of 20 for 4 h, uninfected cells was used as a negative control (Blank). The ratio of cell death was evaluated by the release of LDH in supernatants of infected cells. The activation of caspase-1 (p20) was examined via western blot d. β-actin was blotted as a loading control. Molecular mass markers in kDa are indicated on the right. Original images of immunoblotting were shown in Fig. S2