Fig. 6

HSP60 plays a modulatory role in induction of inflammation in microglia. Knockdown as well as overexpression of HSP60 was done in N9 murine microglial cells by transfection of specific eSiRNA against HSP60 mRNA (Fig. 6a, c) and mouse HSP60 cDNA clone (Fig. 6b, d), respectively, to check subsequent effects on pro-inflammatory factors. a Left upper panel shows representative western blot image of HSP60, iNOS and COX2 in the presence of HSP60 esiRNA (6pM) or scrambled esiRNA and/or IL-1β in N9 microglial cells. Left lower panel shows the bar diagram which represent mean fold change in the levels of HSP60, iNOS, and COX2 with respect to control. One hundred microgram protein was loaded for western blots of iNOS and COX2, and β-actin was used as a loading control. b Right upper panel shows the effect of overexpression of HSP60 on iNOS and COX2 by western blotting. Lower panel shows the bar diagram which represent fold change in the levels of HSP60, iNOS and COX2 with respect to control. One hundred microgram protein was loaded for western blots of iNOS and COX2 and 20 μg for HSP60. β-actin was used as a loading control. The blots are representative of three independent experiments. c, d CBA analysis of pro-inflammatory cytokines TNF-α, MCP-1, and IL-6 in presence of HSP60 esiRNA (c) and mouse HSP60 cDNA clone (d). Data represented are mean ± SD of three independent experiments. *p < 0.05; **p < 0.01 in comparison to control values and ^# p < 0.01 in comparison to IL-1β treatment