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. 2014 Nov 13;63(12):4130–4142. doi: 10.2337/db14-0109

Figure 2.

Figure 2

Expression and phosphorylation of INSR in iPSCs. A: mRNA expression of INSR by qRT-PCR, expressed as percent of maximum value (n = 3 experiments). B: Quantification of INSR protein level from Western blot analysis, expressed as percentage of maximum value (n = 3). C: iPSCs were serum starved for 3 h before 10-min stimulation with 0, 10, or 100 nmol/L insulin. Representative Western blot analysis of INSR and IGF1R expression and phosphorylation are shown (n = 3). D: Quantification of Western blot analysis of P-INSR/IGF1R, represented as percent of maximum value (n = 3). E: iPSCs were serum starved for 3 h before 5-min stimulation with 0 or 100 nmol/L insulin. Immunoprecipitation (IP) of INSR using a β-subunit–specific anti-INSR antibody followed by immunoblotting (IB) with an antiphosphotyrosine antibody (top row) and anti-INSR antibody (bottom row) was performed. Representative blots are shown (n = 2). F: iPSCs were serum starved for 3 h before 5-min stimulation with 0 or 100 nmol/L insulin. IP of IGF1R using a β-subunit–specific anti-IGF1R antibody and IB with an antiphosphotyrosine antibody (top row), anti-P-INSR/IGF1R (middle row), and anti-IGF1R antibody (bottom row) was performed. Representative blots are shown (n = 2). For panels A, B, and D, data are mean ± SEM. *P < 0.05 vs. controls.