Figure S3.

TLR4-SNP macrophages exhibit normal responses to TLR2 and TLR3 agonists, and steady-state Tlr4 mRNA is expressed equivalently in WT and TLR4-SNP macrophages and liver samples. (A) Responses of WT (open bars) vs. TLR4-SNP (gray bars) macrophages to TLR2 (P2C and P3C, 1 µg/ml; 4 h) and TLR3 (p(I:C), 50 µg/ml; 6 h) agonists are not significantly different, in contrast to TLR4 (LPS, 10 ng/ml; 2 h). Results represent the mean ± SEM of four separate experiments and were analyzed by two-way ANOVA with Sidak’s multiple comparison post-test. *, P = 0.046. (B and C) Steady-state levels of Tlr4 mRNA are not significantly different in WT vs. TLR4-SNP macrophage or liver RNA. The data represent Tlr4 mRNA levels from untreated pooled macrophage cultures from the six individual experiments analyzed in Fig. 3 B (B) and livers of 9 WT vs. 10 TLR4-SNP mice (C). qRT-PCR was performed using distinct primer sets for Tlr4 mRNA and two housekeeping genes, Hprt and Actb mRNA, to ensure equivalent RNA loading. Data were analyzed by one-way ANOVA with Sidak multiple comparison post-test. N.S., not significant.