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. 2019 Dec 11;95(10):581–589. doi: 10.2183/pjab.95.039

Figure 3.

Figure 3.

Identification of phosphoamino acids in the β subunit of the insulin receptor in intact cells. Rat hepatoma (Fao) cells were labeled biosynthetically with [32P]orthophosphate and then incubated with or without 1 µM insulin for 15 min, after which the insulin receptor was isolated by immunoprecipitation with specific antibodies followed by SDS-PAGE under reducing conditions. The band corresponding to the 95-kDa immunoreactive protein (β subunit) was excised from the gel, and the protein was eluted by electrophoresis and subjected to hydrolysis with 6 M HCl for 2 h at 110 ℃. The resulting hydrolysate was analyzed by two-dimensional electrophoresis and autoradiography. The positions of spots corresponding to phosphoserine (P-Ser), phosphotyrosine (P-Tyr), and phosphothreonine (P-Thr) are indicated. Modified with permission from ref. 23.