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. 2010 Feb 4;24(3):644–656. doi: 10.1210/me.2009-0357

Figure 1.

Figure 1

Characterization of IGF-IR-floxed primary osteoblasts. A, GH-stimulated JAK2 and STAT5 phosphorylation. Primary osteoblasts were harvested from newborn mice bearing lox-P sites flanking both IGF-IR alleles. Serum-starved cells were treated with vehicle or GH (500 ng/ml) for 15 min. Detergent extracts were resolved by SDS-PAGE and immunoblotted with anti-pTyr JAK2 and anti-pTyr STAT5, respectively. B, Osteoblasts harvested and serum starved as in A were treated with vehicle or GH (500 ng/ml) for 15 min. Detergent extracts were immunoprecipitated with either anti-GHR or nonimmune (NI) serum. Precipitated proteins were resolved by SDS-PAGE and blotted with antiphosphotyrosine. The blots shown are representative of two independent experiments. C, IGF-IR-floxed osteoblasts were infected with either 800 MOI Ad-GFP or 800 MOI Ad-Cre for 48 h. After overnight serum starvation, detergent extracts were resolved by SDS-PAGE and immunoblotted with anti-IGF-IRβ and anti-β-actin, respectively.