Figure 5

NLRP6 is involved in the expression of NLRP3 in P. multocida infected macrophages. Macrophages were infected with the P. multocida at a MOI of 1 for 9 h. Then 100 μg/mL ciprofloxacin was added for an additional 15 h incubation. After 3, 6, 9 and 24 h incubation, cell lysates were collected for RT-PCR and western blot assays. nlrp3 mRNA expression was determined by RT-PCR (A). NLRP3 protein expression was detected by western blot (B) and the ratio of NLRP3 level against GAPDH level was quantified (C). P. multocida-infection after 3, 6, 9 and 24 h detected relative mRNA expression of cxcr2 (D), cxcl1 (E), and cxcl2 (F) were also determined using RT-PCR. The results are representative of three independent experiments. Student t-test was used to analyze the statistical differences for comparisons between two groups. The statistical significance was determined as the p value, *P < 0.05, **P < 0.01, ***P < 0.001, ns: no significance.