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. 2016 Feb 10;291(15):8140–8149. doi: 10.1074/jbc.M115.693598

FIGURE 5.

FIGURE 5.

IGF1 potentiates the association between integrin β3 and IGF1R. A, integrin β3 is recruited to the activated IGF1 receptor. Human fetal osteoblast cells were grown to confluence and serum-deprived for 48 h. IGF1 was then added to a final concentration of 25 ng/ml for 10 min. Immunoprecipitation (IP) revealed increased phosphorylation of IGF1R (pTyr) associated with increased integrin β3 binding. B–D, BiFC and FRET studies. B, expression constructs used in the study. IGF1 treatment increased the interaction between IGF1R and integrin β3 as assessed by BiFC (C) and FRET (D). HEK293 cells were co-transfected with IGF1R and integrin β3 expression vectors. Cells were then treated with 10 ng/ml of IGF1 and BIFC or FRET measurements were made (see “Experimental Procedures”). Association of IGF1R and integrin β3 is indicated by rescue of YFP fluorescence (C) and recovery of CFP fluorescence (D) via transfer of energy from donor IGF1R-YFP fusion protein. For BIFC experiments (C), n = 38 for no IGF1 group; n = 75 for IGF1 group. Values are depicted as the mean ± S.E. *, significant at p ≤ 0.05 by t test.