Fig. 3: Phospho(T25)-cofilin-1 impedes SRF regulation and alters cardiac function in mice. | Nature Communications

Fig. 3: Phospho(T25)-cofilin-1 impedes SRF regulation and alters cardiac function in mice.

From: Actin-microtubule cytoskeletal interplay mediated by MRTF-A/SRF signaling promotes dilated cardiomyopathy caused by LMNA mutations

Fig. 3

a Schematic representation of the luciferase assay for assessing SRF activity. b Bar graph showing quantification of luciferase activity in C2-WT and C2-H222P cells overexpressing SRF response element and transfected or not (NT) with plasmids expressing different mutated cofilin-1 proteins. Results after treatment with selumetinib are also shown (n = 3 independent experiments, mean ± SD). c mRNA expression of SRF target-genes: Acta2, Fhl2, Tnnc, Myom1, Svil, and Msn in C2-H222P and C2-WT cells (n = 6 independent experiments, mean ± SD). d mRNA expression of Srf, Acta2 and Fhl2 in hearts of 3 and 6-month-old male Lmnap.H222P/H222P (H222P) and WT mice (n = 3 mice per condition, mean ± SD). e Schematic representation of methodological protocol. WT mice were injected with AAV vectors encoding either WT or nonphosphorylable cofilin-1(T25A) at 3 months of age. Biochemistry, echocardiography, and transcriptomic analysis were performed at 6 months of age. Subsequent data showed in f–i, are obtained from the hearts of 6-month-old male Lmna WT mice transduced with AAV vectors encoding cofilin-1 or cofilin-1(T25A) and Lmna H222P mice (n = 3 mice per condition, mean ± SD). f Immunoblots showing cofilin-1 (top) and monomeric G-actin (G) and filamentous F-actin (F) (bottom) expression. GAPDH was used as the loading control. Lmna H222P mice are shown as controls, and UT indicates untransduced. g Bar graph showing the left ventricular fraction shortening (FS) values. h Principal component analysis (PCA) of the Affymetrix probe sets. i Unsupervised hierarchical clustering of the Affymetrix probe set results. Statistics: for b and g, one-way ANOVA followed by Tukey’s multiple comparison test; for c and d, unpaired two-tailed t-test. Source data are provided as a Source Data file. For a and e, parts of the figure were drawn by using pictures from Servier Medical Art. Servier Medical Art by Servier is licensed under a Creative Commons Attribution 3.0 Unported License (https://creativecommons.org/licenses/by/3.0/).

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