ILK Induces Hsc70 and Hsp70 Expression in Human Cardiomyocytes.

<p><b>A</b>, (top panels) Phase contrast and fluorescent microscopy images of human IPSC-derived cardiomyocytes (Hu iPS-CMs) (on the left) and primary rabbit cardiomyocytes (on the right) transduced with GFP-linked vector (ad-GFP) versus non transduced cells (C). The Hu iPS-CM cells express monomeric red fluorescent protein and blasticidin resistance under the control of the α-myosin heavy chain promoter that allows simultaneous cardiomyocyte purification and identification. Green fluorescence indicates efficient transduction of ILK constructs. Scale bar, 80 µm. (bottom panels) Western blots showing expression levels of ILK, Hsp70 and Hsc70 in Hu iPS-CMs (on the left) and in primary rabbit cardiomyocytes (on the right) transduced with ILK^<i>WT</i> and ILK^<i>R211A</i> as compared to non-transduced cells and cells transduced with the vector bearing GFP only. GAPDH was used as a loading control. <b>B</b>, Immunoblot for ILK expression levels in Hu IPS-CMs transduced with ad-ILK^<i>WT</i> and ad-ILK^<i>R211A</i> as compared to non-transduced cells following treatment with cycloheximide (CHX) at 80 µM for 0, 3, 6 and 18 hours. <b>C</b>, Western blot analysis showing the protein levels of ILK, Hsp70 and Hsp-90 in Hu iPS-CMs transduced with ad-GFP, ad-ILK^<i>WT</i>, ad-ILK^<i>R211A</i> and in non-transduced cells (C) following treatment with indicated doses of Hsc/p70-ATPase inhibitor (Ver-155008) for 24 hours. GAPDH was used as a loading control. Each experiment was performed at least three times on independent samples and one representative blot is shown.