Fig. 4
From: Myocardin regulates mitochondrial calcium homeostasis and prevents permeability transition
Myocardin regulates mitochondrial calcium homeostasis. a H9c2 cells were transfected with a SR-targeted (above) or mitochondrial (mito)-targeted (below) calcium biosensor. Cells were pre-treated with 2APB (2 µM, 2 h) followed by co-treatment with Forskolin (FSK, 10 µM) and 3-isobutyl-1-methylxanthine (IBMX, 500uM; FSK-I, 18 h), or DMSO as a control vehicle. b Quantification of SR-calcium and c mito-calcium from a. Fluorescence signal normalized to cell area (relative fluorescence) was quantified in 10 random fields. d H9c2 cardiac myocytes were transfected with Myocd or an empty vector, and a SR-targeted calcium biosensor or (e) a mito-targeted calcium biosensor; treated with FSK-I, and quantified. f H9c2 cardiac myocytes transfected with Myocd or an empty vector and treated with FSK-I; stained with TMRM, or (g) calcein-AM and CoCl2, or (h) Mito-Sox to evaluate mitochondrial super oxide, or (i) calcein-AM and Eth HD-1; 10 random fields were quantified for each end-point. j H9c2 cells were transfected with Myocd or empty vector. Protein extracts analyzed as indicated. k H9c2 cells were transfected with sh-Myocd or scrambled control, with the mito-calcium biosensor and stained with Hoechst to visualize nuclei. l Quantification of k. Data are expressed as mean ± SE. *p < 0.05 compared to control, **p < 0.05 compared to treatment, determined by one-way ANOVA
