From: IGF-1 Regulates the Extracellular Level of Active MMP-2 and Promotes Müller Glial Cell Motility Invest. Ophthalmol. Vis. Sci.. 2015;56(11):6948-6960. doi:10.1167/iovs.15-17496 Figure Legend: IGF-1 induces IGF-1R autophosphorylation and downstream activation of intracellular signaling pathways in MIO-M1 cells. (A) Western blot analysis of phosphorylated IGF-1R (p-IGF-1R) in MIO-M1 cells cultured in the presence of 10 nM IGF-1 at indicated times. IGF-1R is shown as a loading control. (B) Representative Western blot assay of phosphorylated AKT (p-AKT) in MIO-M1 cells cultured in the presence of 10 nM IGF-1 at indicated times. AKT 1/2/3 is shown as a loading control. The bars represent the densitometric analysis of the immunoreactive protein expressed as relative intensity calculated from the densitometric intensity of p-AKT signal normalized to AKT 1/2/3. Bars represent the mean ± SD of three independent experiments. The asterisks show statistical differences with respect to control (0 minutes) for P < 0.01 (**) and P < 0.001 (***), respectively. (C) Representative Western blot assay of phosphorylated ERK 1/2 (p-ERK 1/2) in MIO-M1 cells cultured in the presence of 10 nM IGF-1 at indicated times. ERK 1/2 is shown as a loading control. The bars represent the densitometric analysis of the immunoreactive protein expressed as relative intensity calculated from the densitometric intensity of p-ERK 1/2 signal normalized to ERK 1/2. Bars represent the mean ± SD of three independent experiments. ns, nonsignificant. The asterisks show statistical differences with respect to control (0 minutes) for P < 0.001 (***). (D) Western blot assay of lysates from cells preincubated (or not) with the αIR3 antibody in the presence or absence of 10 nM IGF-1, showing p-IGF-1R, p-AKT, and p-ERK 1/2 levels. IGF-1R is shown as a loading control. Date of download: 11/22/2017 The Association for Research in Vision and Ophthalmology Copyright © 2017. All rights reserved.