Figure 4.

Effect of insulin on tyrosine phosphorylation of FAK, paxillin, and Src in HTC cells. (A) Confluent HTC-WT and HTC-IR cells were serum starved for 3 h and treated without or with insulin (100 nM) for times indicated to coincide with a total serum starvation time of 3 h. Cell lysates were prepared and immunoprecipitated (IP) with anti-FAK (α-FAK) and immunoblotted (IB) with α-FAK. Alternatively, cell lysates were immunoprecipitated with anti-PY antibody (α-PY) and immunoblotted with anti-FAK (α-FAK), anti-paxillin (α-PAX), or anti-Src (α-Src), as outlined in MATERIALS AND METHODS. (B) The ECL-developed films were quantitated by scanning densitometry, and the optical absorbance is reported relative to that in untreated samples. Shown is one experiment representative of three.