Fig. 3. KMS99220 inhibits activation of NFκB signaling via HO-1 induction. (A) BV2 cells were treated with 10 µM KMS99220 for 1 h, and then treated with 0.2 µg/ml LPS for additional 1 h. Nuclear extracts were isolated and western blot analysis was performed against p65 NFκB. (B) BV2 cells were treated with 10 µM KMS99220 for 1 h and then exposed to 0.2 µg/ml LPS for additional 0.5 h. Cell lysates were analyzed by western blotting with antibodies to p-IκB or IκB. (C, D) BV2 cells were transfected with siRNA against control or HO-1 for 24 h. (C) Western blot against HO-1 was performed to confirm the knockdown. (D) After treatment with 10 µM KMS99220 for 1 h, the HO-1 knockdown cells were incubated with 0.2 µg/ml LPS for additional 0.5 h and western blot analysis was performed against p-IκB. (E) After pretreatment with SnPP for 1 h, BV2 cells were treated with 10 µM KMS99220 for 1 h followed by 0.2 µg/ml LPS for 0.5 h. Western blot analysis was performed against p-IκB. For quantitation, values obtained from densitometry were normalized against respective loading controls (Lamin B or β-actin) and expressed as fold of respective LPS-treated control (A, B, D, and E) or control siRNA-transfected (C).
© Exp Neurobiol
