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. 2014 Jul 25;155(10):3956–3969. doi: 10.1210/en.2013-2095

Figure 3.

Figure 3.

Effect of inhibitors and Epac siRNA on ghrelin-stimulated SOCS3 expression. A, Representative immunoblot shows that ghrelin (20nM, Ghr) caused a significant (2.59 ± 0.29–fold) increase in SOCS3 expression, which was significantly inhibited (> 80%) by SQ 22536 (100μM). SQ 22536 alone had no significant effect on SOCS3 expression. B, Ghrelin (20nM) caused a 2.26 ± 0.37–fold increase in SOCS3 expression in cells transfected with control siRNA. Ghrelin stimulated SOCS3 expression, which was significantly inhibited (> 90%) by Epac gene silencing. C, Western blot analysis shows that Epac1 expression was significantly inhibited by Epac siRNA (> 60%). D, Effect of PI3K and MAPK inhibitors on ghrelin-stimulated SOCS3 expression. A representative immunoblot shows that ghrelin (20nM, Ghr) caused a significant (2.6 ± 0.21–fold) increase in SOCS3 expression, which was inhibited by > 80% by adenylate cyclase inhibitor SQ 22536 (100μM). The PI3K inhibitor wortmannin (1μM) and the MAPK/MEK inhibitors PD 98059 (30μM) and U0126 (10μM) did not inhibit ghrelin-stimulated SOCS3 expression. E, PKA inhibitor RpcAMP (10μM) did not inhibit ghrelin-stimulated SOCS3 expression. The data are representative of five independent experiments. *, P < .05, significantly different from unstimulated control; **, P < .05, significantly different from ghrelin-stimulated SOCS3 expression.