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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 6. Characterization of a Src-containing Mechano-sensitive Complex in Osteoblasts.

Figure 6

A. and B. Western blots of detergent-insoluble fractions isolated from static MC3T3 cells and from cells exposed for 5 min to orbital fluid shear stress (FSS; 120 rpm) (A). The bar graphs in B summarize three separate experiments (mean ± SEM; n=3; *p < 0.05 for static versus shear-stressed cells).

C. and D. PKG II (white) and Shp-2 (red), or Src (white) and Shp-2 (red) colocalization with vinculin (green) was examined in static and shear-stressed (FSS) MC3T3 cells (C). Focal adhesions were defined as vinculin-positive membrane complexes > 0.5 μm size, and colocalization of PKG II and Shp-2, or Src and Shp-2 with focal adhesions was scored as a percentage of total focal adhesions (D shows the mean ± standard error of proportion for three experiments, with ~35 cells evaluated per condition; *p < 0.05 for static versus shear-stressed cells; bar 5 μm).

D. Colocalization of αIIb/β3 integrins with PKG II monitored by bimolecular fluorescence complementation (BiFC) between αIIb-VC and wild-type PKG II-VN (VN-WT) or the membrane binding-deficient G2A mutant PKG II-VN (VN-G2A) in MC3T3 cells. All cells were transfected with human β3 and αIIb-VC (representative confocal images of three separate experiments; bar 5 μm).