Fig. 6.

JNK functions upstream of HSP70 in regulation of TNFα production after heat and LPS. WT or ANXA1^−/− macrophages were incubated at either 37 or 42 °C for 1 h prior to stimulation with LPS (100 ng/ml). Cells were lysed for a Western blotting for HSP70 after 30 and 60 min of LPS or b real-time PCR for HSP70 mRNA after 30 min of LPS. c Cells were pretreated for 1 h with the HSP70 inhibitor VER155008 prior to heat and LPS treatment. Cells were lysed for Western blotting for the indicated proteins after 30 min of LPS, and d TNFα production was measured by ELISA after 24 h. e Cells were pretreated for 1 h with the JNK inhibitor SP600125 (10 μM) prior to heat and LPS treatment. Cells were lysed for Western blotting for the indicated proteins after 30 min of LPS, and d TNFα production was measured by ELISA after 24 h. *p < 0.05, **p < 0.01 vs respective LPS controls