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. 2011 Sep 20;2:30. doi: 10.3389/fendo.2011.00030

Figure 3.

Figure 3

Effects of pathway blockade on growth hormone-induced activation of (A) ERK, (B) Akt, and (C) STAT5. CHO-K1 cells were stably transfected with cDNA encoding rainbow trout GHR1 and GHR2. Transfected cells were pretreated with or without specific inhibitors [10 μM of the MEK inhibitor, U0126; 20 μM of the PI3K inhibitor, LY294002 (LY); 25 μM of the Akt inhibitor, 1L6-hydroxyymethyl-chiro-inositol-2-(R)-2-O-methyl-3-O-octadecyl-sn-glycerocarbonate (Carb); 200 μM of the STAT5 inhibitor, N′-((4-oxo-4H-chroen-3-yl)methylene)nicotinohydrazide (Nico); and 50 μM of the JAK2 inhibitor, 1,2,3,4,5,6-hexabromocyclohexane (Hex)] for 2 h, then treated with or without 100 ng/ml growth hormone (GH) for 10 min (control is 0 ng/ml GH); after which time, the cells were collected and lysed. Cell lysates were separated by SDS-PAGE followed by Western immunoblotting using phospho-specific (pERK1/2, pAkt, or pSTAT) and total (tERK1/2, tAkt, or tSTAT) antibodies. Data are expressed as percentage of control and are presented as means ± SEM (n = 8). For a given GHR subtype, groups with different letters are significantly different from each other (P < 0.05); * designates a significant difference (P < 0.05) between subtypes within a given treatment.