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. Author manuscript; available in PMC: 2011 Dec 21.
Published in final edited form as: Sci Signal. 2010 Dec 21;3(153):ra91. doi: 10.1126/scisignal.2001423

Figure 3. Fluid Shear Stress- and cGMP-induced Src Activation Mediated by Shp-1/2.

Figure 3

A. MC3T3 cells were treated with 10 μM vanadate for 1 hour prior to a 5 min exposure to fluid shear stress (FSS; 12 dynes/cm2) or 100 μM 8-CPT-cGMP (cGMP). Src phosphorylation was analyzed as in Fig. 1F (mean ± SEM; n=3; p < 0.05 for vanadate versus control).

B. hPOBs were treated with vanadate and cGMP as in A (images are representative of three experiments)

C. and D. MC3T3 cells were transfected with siRNAs targeting GFP, Shp-1, Shp-2, RPTP-α, or PTP-1B, and phosphatase expression was quantified by Western blotting (whole cell lysates for Shp-1, -2, and PTP-1B, and membrane lysates for RPTP-α). In D, cells were exposed to fluid shear stress or treated with 100 μM cGMP for 5 min and Src phosphorylation was analyzed as in Fig. 1F (mean ± SEM; n=3; p < 0.05 for the comparison between siRNAs targeting Shp-1 or Shp-2 versus GFP).

E. and F. MC3T3 cells were transfected with siRNAs targeting GFP, Shp-1 (E) or Shp-2 (F). Cells were infected with adenovirus encoding LacZ, siRNA-resistant human Shp-1 (E), or Shp-2 (F), and received 100 μM cGMP for 5 min(representative of three experiments).