QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 6. PGC-1, NRF1 and TFAM induction in cardiomyocytes. (A) PGC-1 associates with NRF1 in H9c2 cells after treatment with CO. Equal protein was immunoprecipitated (IP) with anti-PGC-1 or IgG (control), separated by SDS-PAGE, and blotted with anti-NRF1. After a 6-hour treatment with CO (100 mM DCM), PGC-1 partners with NRF1 and is abrogated by PI3-K/Akt inhibition (LY29). Loading was evaluated with 10% of the final immunoprecipitate on gels stained with Coomassie Blue. Data are representative of n=3. (B) ChIP assay for NRF1 and NRF2 binding to the Tfam promoter in H9c2 cells. Input lanes show PCR product prior to immunoprecipitation (loading control). DNA was analyzed by PCR with specific primers for Tfam promoter. CO (DCM 100 µM) increased binding of both transcription factors to Tfam promoter. After mtCAT transfection, CO had no significant effect on NRF1. (C) Mitochondrial BrdU incorporation after CO in cardiomyocytes. Confocal images of MitoTracker Green and Cy3 immunostaining for BrdU were obtained by BrdU labeling (red) as indicated followed by double-label confocal microscopy. Control H9c2 cells (A-C); cells exposed to CO (DCM 100 µM) for 6h followed by washing and 6h incubation (D-F). Panels A and D show mitochondria with MitoTracker (A,D); Red Cy3 immunostaining for BrdU (B,E). merged images of MitoTracker and Red Cy3 (C,F). Cytochrome P₄₅₀ inhibitor (SKF-525A) abrogated CO-induced BrdU incorporation. Yellow/orange signal (G-I) indicates mitochondrial BrdU localization.

Return to article