IL-11 promotes Ang II-induced autophagy inhibition and mitochondrial dysfunction in atrial fibroblasts

Figures

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  Figure 1. Identification of the isolated atrial fibroblasts. (a) The cellular morphology of isolated atrial fibroblasts was captured under an inverted microscope. (b) The expression of vimentin, vWF, and Desmin of isolated atrial fibroblasts was analyzed by IF.

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  Figure 2. IL-11 enhances phenotypic changes in Ang-II-elicited atrial fibroblasts. Atrial fibroblasts were stimulated with 1 μM Ang-II for 24 h. IL-11 sequences were inserted into pcDNA vector plasmids and then transfected into atrial fibroblasts to overexpress IL-11. (A) IL-11 protein expression was detected by western blotting. Data were analyzed after normalization with GAPDH. **P < 0.01 vs. Control; ##P < 0.01 vs. Ang-II + Vector. N = 3. (B) α-SMA level was determined by immunofluorescence assays. Scale bar = 100 µm.

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  Figure 3. IL-11 increases ROS production in Ang II-induced atrial fibroblasts. ROS levels were examined after atrial fibroblasts were incubated with DHE for 30 min at 37°C. Scale bar = 200 µm.

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  Figure 4. IL-11 reinforces mitochondrial dysfunction in Ang II-induced atrial fibroblasts. (a) MMP levels were examined after atrial fibroblasts were stained with JC-1. Scale bar = 100 µm. (b and c) Relative protein expression of PGC-1α and TFAM was detected by western blotting. Data were analyzed after normalization with GAPDH. ***P < 0.001 vs control; #P < 0.05 and ###P < 0.001 vs Ang-II + vector. N = 3.

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  Figure 5. IL-11 facilitates autophagy inhibition in Ang II-elicited atrial fibroblasts. Relative protein expressions of LC3 I, LC3 II, LC3 II/I, p62, and Beclin-1 were determined by western blotting. Data were analyzed after normalization with GAPDH. **P < 0.01 and ***P < 0.001 vs control; ##P < 0.01 and ###P < 0.001 vs Ang-II + vector. N = 3.

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  Figure 6. IL-11 enhances mTOR pathway activation in Ang-II-evoked atrial fibroblasts. Relative protein expressions of mTOR, p-mTOR, and p-mTOR/mTOR were quantified by western blotting. Data were analyzed after normalization with GAPDH. *P < 0.05 vs control; ###P < 0.01 and ###P < 0.001 vs Ang-II + vector. N = 3.