Skip to main content
. 2015 Aug 19;10(8):e0136175. doi: 10.1371/journal.pone.0136175

Fig 2. Confirmation of TLR4 expression in RF/6A cells and the effects of TLR4 knockdown on LPS-induced CXCR4 and CXCR7 expression.

Fig 2

(A) RF/6A cells were stimulated by LPS (1μ/ml) and were compared for the expression of TLR4 with that of untreated cells. After 24 h, the amounts of TLR4 mRNA and protein were quantified by real-time RT-PCR and western blot. In addition, immunofluorescence microscopic analysis was performed for the identification of TLR4 location and expression. TLR4 was labeled by green fluorescence (FITC), and the nuclei were counterstained with DAPI (blue). (B) RF/6A cells transfected with the TLR4 siRNA sequence exhibited a marked reduction in TLR4 mRNA and protein levels, compared with negative control sequence (control). (C) After LPS treatment, western blot and real-time RT-PCR analyses shown that knockdown of TLR4 inhibited LPS-mediated CXCR4 and CXCR7 expression. Data are shown as the mean ± SD of three separate experiments. Statistical significance determined using Student’s t test or one-way ANOVA with *P < 0.05, **P < 0.01.