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. 2019 Jun 3;9:8228. doi: 10.1038/s41598-019-44635-z

Figure 1.

Figure 1

Acute and chronic decreases in synaptic insulin responsiveness. Ex vivo insulin responsiveness assays were conducted on isolated hippocampal synaptosomes at various time-points after traumatic brain injury. Representative Wes analysis of insulin-stimulated and unstimulated synaptosomes isolated from the ipsilateral hippocampus (a) at 2 DPI (n = 4 for both sham and TBI), 7 DPI (n = 4 sham, n = 6 TBI), 1 MPI (n = 5 sham, n = 7 TBI), and 3 MPI (n = 4 for both sham and TBI) animals. Quantitative graph of Wes analysis showing (b) the ratio of insulin-stimulated P-IR/β-tubulin to IR/β-tubulin demonstrating that synaptosomal insulin responsiveness is chronically decreased after TBI, (c) IR/β-tubulin showing the changes in insulin receptor level at the synapse, and (d) the ratio of unstimulated P-IR/β-tubulin to IR/β-tubulin as an indication of basal phosphorylation levels of the receptors. Representative Wes analysis of insulin-stimulated and unstimulated synaptosomes isolated from the contralateral hippocampus (e) from 2 DPI (n = 3 sham, n = 4 TBI), 7 DPI (n = 3 sham, n = 5 TBI), 1 MPI (n = 5 sham, n = 4 TBI), and 3 MPI (n = 3 sham, n = 4 TBI) animals with corresponding quantitative graphs of (f) the ratio of insulin-stimulated P-IR/β-tubulin to IR/β-tubulin, (g) IR/β-tubulin, and (h) the ratio of unstimulated P-IR/β-tubulin to IR/β-tubulin. A Wes assay was run with samples probed for P-IR and β-tubulin as a loading control. Samples were then run through a second Wes assay and probed for total IR and β-tubulin. Full length Wes images are presented in Supplementary Figs S1 and S2. Statistical significance was determined by unpaired t-test analysis. Error bars represent standard error. *p < 0.05; **p < 0.01.