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Figure 4 | Cell Communication and Signaling

Figure 4

From: Up-regulation of COX-2/PGE2 by endothelin-1 via MAPK-dependent NF-κB pathway in mouse brain microvascular endothelial cells

Figure 4

ET-1-induced COX-2 expression is mediated through MAPKs phosphorylation. (A, B) Cells were treated with 10 nM ET-1 for (A) 6 h and (B) 1 h in the absence or presence of U0126, SB202190, or SP600125. The COX-2 protein and mRNA expression were determined by Western blot and RT-PCR. (C) Time dependence of ET-1-stimulated ERK1/2, p38 MAPK, and JNK1/2 phosphorylation, cells were incubated with 10 nM ET-1 for the indicated times in the absence or presence of U0126 (1 μM), SB202190 (300 nM), or SP600125 (300 nM). (D) Cells were transfected with siRNA of ERK2, p38 MAPK, or JNK1 and then exposed to ET-1 for 6 h. (E) Cells were pretreated with BQ-788 (1 μM), GPA2 (1 μM), or GPA2A (1 μM) for 1 h and then incubated with ET-1 (10 nM) for the indicated times. The cell lysates were collected and analyzed by Western blotting using an anti-COX-2, anti-phospho-ERK1/2, anti-phospho-p38 MAPK, anti-phospho-JNK1/2, anti-ERK2, anti-p38 MAPK, anti-JNK1, or anti-GAPDH (as an internal control) antibody. Data are expressed as mean ± SEM of at least three individual experiments (n=3 in each group; *P<0.05, #P<0.01 as compared with ET-1 alone).

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