Figure 7

MAP kinase is activated by SDF1 and ERK inhibitor reduces MMP1 production and CS cell invasion. JJ cells were cultured with 10 ng/ml SDF1 or pretreated for 2 h with 10 μmol/L U0126, 40 nM SP600125, or 100 nM SB203580 in hypoxia. A, Whole cell lysates at indicated time points were subjected to Western blot analysis for total and phophorylated ERK, JNK, and p38. B, Cell lysates were used for Western blot analysis of total MMP-1 after 48 h, SDF1 10 ng/ml, AMD3100 0.1 μM, or U0126 10 μmol/L in hypoxia. C, JJ cells were cultured for 24 h in hypoxia with DMSO (Mock), SDF1 10 ng/ml alone or after 2 h pretreatment with MAP kinase inhibitors (U0126 10 μM, SP600125 40 nM, SB203580 2 μM), then invasion assay was performed for 72 h in hypoxia with SDF1 50 ng/ml in lower chamber only and inhibitors in both chambers. *, p < 0.01, compared to Mock; **, p < 0.05, compared to SDF-1. Bar graphs represent data from three independent experiments. D, representative photographs of stained cells on the lower side of matrigel membrane from one of three experiments in C. Original magnification 100×.