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. Author manuscript; available in PMC: 2017 Feb 1.
Published in final edited form as: Biochim Biophys Acta. 2015 Dec 2;1862(2):240–251. doi: 10.1016/j.bbadis.2015.12.004

Figure 6. MiR-let-7g regulates PI3K signaling to modulate AKT activation by targeting PIK3IP1.

Figure 6

A. PIK3IP1 is induced by hypoxia in NRVMs. NRVMs were cultured in normoxia and hypoxia and mRNA levels of PIK3IP1 were measured using RT-PCR (n=4). B. miR-let-7g modulates mRNA levels of PIK3IP1. NRVMs were cultured in normoxia and transduced with lenti-let-7g and NT vector and mRNA levels of Lin28 and PIK3IP1 were measured using RT-PCR (n=4). C. miR-let-7g targets PIK3IP1 to cardiac myocyte RISC. NRVMs were cultured in normoxia and transduced with lenti-let-7g and NT vector so as to program the myocyte RISC. Following immunoprecipitation with Ago2 antibody as described in Methods levels of various miRs and genes were quantified in the immunoprecipitated RNA fraction using RT-PCR. (top) Transcript levels of miR-let-7g, miR-142 and snoRNA. (bottom) mRNA levels of PIK3IP1, Lin28, β-actin and 18S. Transcript levels were normalized to immunoprecipitated AGO2 protein (n=3). D. miR-let-7g functionally targets WT PIK3IP1 3′fUTR. 293Ts expressing miR-let-7g and NT were transfected with pGLO dual luciferase vector. Renilla luciferase units were normalized to firefly luciferase units (n=5).