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. 2020 Mar 20;69(6):1193–1205. doi: 10.2337/db19-0944

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© 2020 by the American Diabetes Association

Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. More information is available at https://www.diabetesjournals.org/content/license.

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Analysis of background mechanism underlying IRS downregulation in CD36-overexpressing INS-1 cells. A: mRNA expression levels of Irs1 and Irs2. The expression levels were normalized to Ppia and Hprt. N = 6 for each group. B: Phosphorylation levels of IRS1 Ser307 (pSer307) and Ser612 (pSer612). Phosphorylated protein levels were normalized to the total protein levels. N = 4 for each group. Western blot analysis of cytoplasmic (Cyt) and membrane (Mem) protein levels of PKCδ (C) and PKCε (D). N = 3 for each group. The total protein images obtained using the Stain-Free technology are shown as a loading control (Loading). DOX− and DOX+ represent INS-1 cells without and with doxycycline-induced CD36 overexpression, respectively. The P values were determined by Student two-tailed t test (unpaired). *P < 0.05; **P < 0.01. N.D., not determined.