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. 2021 Feb 11;12:593862. doi: 10.3389/fphys.2021.593862

FIGURE 1.

FIGURE 1

Impact of LRP1 on CTRP9-induced mitochondrial biogenesis. H9C2 cardiomyoblasts were transfected with siRNA directed against rat LRP1 or mock siRNA for 48 h and subsequently treated with CTRP9 (4 μg/ml) for the indicated times. (A) Cells were treated with CTRP9 (4 μg/ml) for 20 min and afterward analyzed by Western Blotting for p-AMPK, total-AMPK and LRP1. Representative Western blots and densitometry of protein data are shown. GAPDH served as a loading control. (B) Cells were treated with CTRP9 (4 μg/ml) for 24 h. mRNA expression of ND1 and Cox I as determined by real-time PCR (left panels) and representative Western blots for ND1, Cox I, LRP1, and GAPDH (right panel) are shown. (C) Cells were treated with CTRP9 (4 μg/ml) for 24 h. Genomic DNA was isolated from the cells and afterward analyzed by Real-time PCR for the expression of a mitochondrial transcript and a nuclear transcript in order to estimate mtDNA content (left panel). Citrate synthase (CS) activity (right panel) was measured in H9C2 cell lysates. All values were normalized to total protein content of the respective sample. All data are mean ± SEM, 4 independent experiments, n = 8 per group, *p < 0.05, **p < 0.01.