Figure 3.

Blocking MD2 inhibits NF-κB and inflammatory responses in NEC rats. (A, B) Immunoprecipitation assays showed that MD2 inhibitor pretreatment significantly reduced the formation of the TLR4–MD2 complex in rat intestinal tissue. IB indicates immunoblotting antibody, and IP indicates precipitating antibody (n=3). (C, D) Western blot analysis of TLR4 signaling-related proteins, NF-κB activation and nuclear translocation of the p65 subunit of NF-κB (n=3). (E, F) TNF-α and IL-6 levels in intestinal tissue lysates were detected by ELISA, and values were normalized to the total protein (n=8). The p value was analyzed by one-way ANOVA with multiple comparisons, followed by Tukey’s post hoc test. *p < 0.05, **p < 0.01, ***p < 0.001.