Fig. 10

Role of MEK3/6 in HSP60-induced inflammation. N9 cells were transfected with 4 μg of HSP60 cDNA clone and/or, MEK3/6 specific eSiRNA (6pM) for 24 h, and the effect on inflammation was assessed by western blot (a) and cytokine bead array (b). a Western blot analysis of phospho-MEK3/6 and total MEK3/6, phospho- and total p38, iNOS and COX2 after inhibition of MEK3/6 and overexpression of HSP60. Blots are representative of three different experiments with similar results. One hundred microgram protein was loaded for western blots, and β-actin was used as a loading control. Graphs represent the mean fold change in the phosphorylation of ERK1/2, JNK, and p38 with respect to their respective total proteins and represents mean fold change in the expression of iNOS and COX2 with respect to control. b Effect of knockdown of MEK3/6 on pro-inflammatory cytokines. CBA analysis of TNF-α, MCP-1, and IL-6 after transfection of N9 microglial cells with HSP60 cDNA clone and/or, MEK3/6 eSiRNA (i–iii) Data represented are mean ± SD of three independent experiments. *p < 0.05; **p < 0.01 in comparison to control values