FIG. 4.

ERK binding to α₉β₁ integrin and FAK is downregulated by hUCBSC treatment. (A) Around 800 μg of total soluble protein were taken from U251 and 5310 control cells and shERK-transfected samples. The protein samples were incubated with 2 μg/μL of ERK antibody. The mixture containing total soluble protein and respective antibody was incubated with 50 μL of protein A/G agarose beads for 30 min in ice. The ERK immunoprecipitated samples were immunoblotted against FAK and integrin α₉β₁. (B) Eight hundred microgram of total soluble protein from U251 and 5310 control and hUCBSC-treated tissue lysates were incubated with 2 μg/μL of ERK antibody. The ERK-immunoprecipitated samples were immunoblotted against FAK and integrin α₉β₁. The same blots were stripped and re-probed with anti-ERK antibody. (C) Eight hundred microgram of total soluble protein from U251 and 5310 control and hUCBSC-treated tissue lysates were immunoprecipitated with c-Myc, and the samples were immunoblotted against pERK, MAX, and FAK. The same blots were stripped and re-probed with anti-c-Myc antibody. (D) U251 and 5310 control and hUCBSC-treated tissue lysates were immunoprecipitated with FAK and immunoblotted against c-Myc. The same blot was stripped and re-probed with anti-FAK antibody. These data are representative of at least 2 individual experiments.