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. Author manuscript; available in PMC: 2012 Nov 1.
Published in final edited form as: Cell Signal. 2011 Jun 25;23(11):1794–1805. doi: 10.1016/j.cellsig.2011.06.014

Figure 3.

Figure 3

A–B. The effects of SFK inhibition on the activation of downstream effectors of PRL-R Serum-starved T47D (A) or MCF-7 (B) cells were either left untreated (−) or were treated (+) with Su6656 (10 μM, 30 min) before stimulation with 10 nM PRL for the indicated time intervals. Phosphorylated forms of JAK2 (Tyr Tyr1007/1008), STAT5 (Tyr694), FAK (Tyr925), Gab1 (Tyr627), SHP2 (Tyr542), Akt (Ser473), MEK1/2 (Ser217/Ser221) and ERK1/2 (Thr202/Tyr204) proteins were detected by IB of the TCL with Abs against respective antigens. GAPDH levels were detected with anti-GAPDH Abs and used as protein loading control. Representative blots are shown (n=3). B. Phosphorylated STAT5 (white bars), Akt (dark grey bars) and ERK1/2 (light grey bars) in TCL of Su6656-treated cells were quantified by densitometry and are expressed as percent of control of the corresponding phospho-protein measured in TCL of PRL-stimulated cells incubated without Su6656. Data represent the mean ± SD of three independent experiments. C–D. The effects of FAK inhibition on the activation of downstream effectors of PRL-R. Serum-starved T47D cells were either left untreated (−) or were treated (+) with PF573228 (0.5 μM, 2 h) before stimulation with 10 nM PRL for the indicated time intervals. Phosphorylated forms of FAK (Tyr397), FAK (Tyr576/577), FAK (Tyr925), SFKs (Tyr416), STAT5 (Tyr694), Akt (Ser473), MEK1/2 (Ser217/Ser221) and ERK1/2 (Thr202/Tyr204) proteins were detected by IB of the TCL with Abs against respective antigens. GAPDH levels were used as protein loading control. Representative blots are shown (n=3). D. The ratio of phospho-ERK1/2:total ERK1/2 at each time point was expressed as fold changes over basal levels.