Figure 5

The over-expression of acetate assimilation system promoted pro-inflammatory responses of APEC infected macrophages. A Determining the transcriptional differences of pro-inflammatory cytokines (IL-1β, IL-6, IL-8, IL-12β, and TNF-α), anti-inflammatory cytokines (IL-4, IL-10, and IL-13), and iNOS between FY26 infected HD11 cells and non-infectious cells. qRT-PCR data acquired from three independent experiments were used to determine the transcription of selected cytokines genes during wild-type FY26 replication within HD11 cells at 4 hpi and 8 hpi. Significant differences were statistically determined using one-way ANOVA analysis (*P < 0.01), and the mean values ± the standard errors were shown. B Determining the effect of acs-yjcH-actP deletion on proinflammatory responses in RAW264.7 macrophages. Transcriptional differences of IL-1β, IL-6, IL-8, IL-12β, TNF-α, and iNOS among wild-type FY26, FY26Δacs-yjcH-actP, and FY26Cacs-yjcH-actP infected HD11 cells at 4 hpi were analyzed by qRT-PCR relative to that of uninfected HD11 cells. C Expression analysis of nitric oxide (NO) and secreted cytokines in FY26, FY26Δacs-yjcH-actP, and FY26Cacs-yjcH-actP infected RAW264.7 macrophages at 8 hpi. The cultured supernatant of infected cells was harvested at 8 hpi or the uninfected cultured. ELISA assay was conducted to evaluate the concentration of NO and proinflammatory cytokines (IL-1β, IL-6, IL-8, IL-12β, and TNF-α) in the culture supernatant of RAW264.7 macrophages to determine the effect of acs-yjcH-actP deletion on proinflammatory responses in RAW264.7 macrophages. D Expression analysis of nitric oxide (NO) and secreted cytokines in FY26, FY26Δacs-yjcH-actP, and FY26Cacs-yjcH-actP infected RAW264.7 macrophages at 16 hpi. Statistical difference was determined using one-way ANOVA analysis (*, P < 0.01), and the mean values ± the standard errors were shown.