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. Author manuscript; available in PMC: 2012 Dec 1.

Published in final edited form as: Mol Immunol. 2011 Oct 26;49(3):423–432. doi: 10.1016/j.molimm.2011.08.026

A and B: The effect of LPS, pam2CSK4 or LPS plus pam2CSK4 on phosphorylation of p38 MAPK. U937 cells were treated with LPS, pam2CSK4 or both for 0, 5, 10, 20 or 60 min. At each time point, cells were lysed and the cell lysate was subjected to immunoblotting to detect phosphorylated and total p38 MAPK. C: The effect of p38 MAPK inhibitor SB239063 on IL-6 secretion in response to LPS plus pam2CSK4. U937 cells were treated with or without 100 ng/ml of LPS plus pam2CSK4 in the absence or presence of 5 or 10 μM of SB239063 for 24 h. After the treatment, IL-6 in culture medium was quantified using ELISA. D to G: The effect of LPS, pam2CSK4 or LPS plus pam2CSK4 on ERK MAPK, and JNK MAPK. Similar studies as described in A and B were conducted to determine phosphorylated and total ERK and JNK MAPK. The bands for phosphorylated p38, ERK and JNK MAPK in the images were quantified using densitometric scanning and normalized to total p38, ERK and JNK MAPK (B, E and G). The data presented are the representative of 3 independent experiments with similar results. P-p38, P-ERK, P-JNK: phosphorylated p38, phosphorylated ERK, phosphorylated JNK. T-p38, T-ERK, T-JNK: total p38, total ERK, total JNK.

A and B: The effect of LPS, pam2CSK4 or LPS plus pam2CSK4 on phosphorylation of p38 MAPK. U937 cells were treated with LPS, pam2CSK4 or both for 0, 5, 10, 20 or 60 min. At each time point, cells were lysed and the cell lysate was subjected to immunoblotting to detect phosphorylated and total p38 MAPK. C: The effect of p38 MAPK inhibitor SB239063 on IL-6 secretion in response to LPS plus pam2CSK4. U937 cells were treated with or without 100 ng/ml of LPS plus pam2CSK4 in the absence or presence of 5 or 10 μM of SB239063 for 24 h. After the treatment, IL-6 in culture medium was quantified using ELISA. D to G: The effect of LPS, pam2CSK4 or LPS plus pam2CSK4 on ERK MAPK, and JNK MAPK. Similar studies as described in A and B were conducted to determine phosphorylated and total ERK and JNK MAPK. The bands for phosphorylated p38, ERK and JNK MAPK in the images were quantified using densitometric scanning and normalized to total p38, ERK and JNK MAPK (B, E and G). The data presented are the representative of 3 independent experiments with similar results. P-p38, P-ERK, P-JNK: phosphorylated p38, phosphorylated ERK, phosphorylated JNK. T-p38, T-ERK, T-JNK: total p38, total ERK, total JNK.