Fig. 5.

Effect of L. donovani infection on Pam3cys stimulated activation of MAPK P³⁸ in anisomycin and U0126 treated cells. THP-1 cells were infected with L. donovani at 10:1 parasites to cell ratio and simultaneously stimulated with Pam3cys and P³⁸ activator anisomycin (0·1, 1 and 10 ng/ml) or pretreated with ERK1/2 inhibitor U0126 (0·1, 1 and 10 µM) for 1 h followed by addition of L. donovani at 10:1 parasites to cell ratio and Pam3cys. Cell lysates were prepared after incubating for 15 min, subjected to electrophoresis, transferred on nitrocellulose membrane and probed with anti-phosphorylated P³⁸ and non-phosphorylated P³⁸ antibodies. The results are expressed as ratio of densitometric readings of phosphorylated P³⁸ to non-phosphorylated P³⁸ (A). L. donovani infected THP-1 cells were activated with anisomycin and Pam3cys (1 µg/ml) or U0126 pretreated and infected cells were activated with Pam3cys (1 µg/ml). Supernatant were collected after 24 h and IL-12p40 (B) and IL-10 (C) measurement done by ELISA. Mean + SD of three independent experiments is shown and group comparison was done using student's t test P < 0·05 considered significant (*P < 0·05, **P < 0·005).