Figure 8

Silencing of TLR2 down-regulated CP/CP-PGN-induced activation and differentiation of RAW264.7 cells. The expression levels of TLRs were analyzed by qRT-PCR to select out which TLR changed most obviously (a). RAW264.7 cells transfected 24 h with TLR2 siRNA/control siRNA, then the mRNA of TLR2 were assayed by qRT-PCR (b). CP and CP-PGN pretreated RAW264.7 or RAW264.7 transfected with TLR2 siRNA co-stimulated with MRSA to analysis the phagocytosis rate (c). RAW264.7 or RAW264.7 transfected with siTLR2 were co-cultured with CP or CP-PGN for 24 h to detect the production of NO (d), for 3 h to detect cytokines (e,f,g), iNOS (h) and arginase-1 (i) by qRT-PCR. CD86 were determined by flow cytometric analysis (j) and cell migration assay was shown (k). Each graph represents the average of three replications. The significant differences compared with the untreated group were analyzed by Student’s t test, ^* P < 0.05, ^** P < 0.01, ^*** P < 0.001 vs. untreated. CP = C. pyruviciproducens; CP-PGN = PGN of C. pyruviciproducens.