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miR-375 overexpression in expanded islet cells downregulates the PDPK1-AKT pathway.

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posted on 13.04.2015, 06:11 by Gili Nathan, Sharon Kredo-Russo, Tamar Geiger, Ayelet Lenz, Haggai Kaspi, Eran Hornstein, Shimon Efrat

A,B. Immunoblotting of PDPK1 in expanded islet cells at the indicated passages. HSC70 served as loading control. Values are mean±SE (n = 3 donors). C, D. Immunoblotting of PDPK1 in expanded islet cells infected at passages 4–6 with miR-375 or empty viral vectors. Values are mean±SE (n = 3 donors). E. Immunoblotting of PDPK1 in expanded islet cells at passages 4–6 infected with 4 different PDPK1 shRNAs. NT, nontarget. F. Changes in insulin transcript levels in expanded islet cells infected at passages 4–6 with PDPK1 or NT shRNAs, and analyzed 5 days later by qRT-PCR. Values are mean±SE (n = 3 donors), relative to NT. *p<0.05. G,H. Immunoblotting of phosphorylated AKT (Thr308) in expanded islet cells at the indicated passages. Values are mean±SE (n = 3 donors). I, J. Immunoblotting of phosphorylated AKT in expanded islet cells infected at passages 4–6 with miR-375 or empty viral vectors. Values are mean±SE (n = 3 donors). K,L. Immunoblotting of AKT in expanded islet cells infected at passages 4–6 with miR-375 or empty viral vectors. Values are mean±SE (n = 3 donors). *p = 0.006. M. Scheme of miR-375 effect on AKT targets through PDPK1.

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