Figure 5

Tissue specific phosphorylation of Grb14. Retinal, heart, and liver protein lysates from dark- and light-adapted rats were incubated with the GST-vSrc-SH2 fusion protein, followed by GST pull-down assay. The bound proteins were subjected to immunoblotting with anti-Grb14 (A) and anti-GST (C) antibodies. Tissue lysates underwent immunoblot analysis with anti-Grb14 antibody (B). Blood was drawn from control and Ins2^Akita mouse tail veins, and the blood glucose levels were monitored with a glucometer (D). Data are mean ± SE, n = 8, *p < 0.001. Control and Ins2^Akita mouse retinal lysates were subjected to GST-vSrc-SH2 fusion protein, followed by GST pull-down assay. The bound proteins were subjected to immunoblot analysis with anti-Grb14 (E) and anti-GST (F) antibodies. Densitometric analysis of 4 independent immunoblots of Grb14 was performed in the linear range of detection and absolute values were then normalized to v-Src-SH2 (G). Values are mean ± SEM, (n = 4), *p < 0.05. The binding of Grb14 to the v-Src-SH2 domain in control mice was set at 100 percent.