Figure 5. MD2 knockdown reduced Ang II-induced inflammation by inhibition of MAPKs phosphorylation. VSMCs were transfected with siRNA against MD2 for 6 h and then incubated with Ang II for 6 h (panel A) and 30 min (panel B, C). (A) The level of TNF-α and IL-6 were detected using real-time qPCR assay (n = 3; **p<0.01 compared to Ctrl; ##p<0.01 compared to Ang II). (B, C) Expressions of P-ERK, P-JNK, P-P38 in the cultural medium were detected by western blot (n = 3; **p<0.01, ***p<0.001 compared to Ctrl; ##p<0.01 compared to Ang II). (D–I) Representative immunofluorescence staining images and quantification results for P-ERK (green), P-JNK (green), P-P38 (green) in mouse aortas. Tissues were counterstained with DAPI (blue) (scale bar = 50 μm) (n = 10; **p<0.01, ***p<0.001 compared to Vehicle; ###p<0.001 compared to Ang II).