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| Activin A signaling regulates epiMT (A) mRNA expression levels for Activin receptor type 2A (ACVR2A), Activin receptor type 2B (ACVR2B), Activin receptor type 1B (ALK4), Inhibin subunit beta A (INHBA) and Inhibin subunit beta B (INHBB) determined in cobble fetal and adult EPDCs cultured for 3 h after removal of SB (n 6). Raw Ct values per condition are shown in italics. * p < 0.05, ns not significant (unpaired Student's t-test). Data are displayed as mean + SEM. (B) Immunofluorescent staining for phalloidin and αSMA in fetal EPDCs cultured for 5 days in control (CTRL) or FST containing medium. Orange arrows indicate examples of epithelial cobblestone-shaped cells, blue arrows of mesenchymal spindle-shaped cells. Scale bar: 100 µm. (C) Quantification of αSMA positive surface area of FST treated cells relative to CTRL (n 7). * p < 0.05 (paired Student's T-Test). (D) Phalloidin staining in adult EPDCs cultured for 5 days in control medium (CTRL), medium containing Activin A, or Activin A in combination with SB431542. Scale bar: 100 µm. (E) mRNA expression levels of WT1, POSTN, CDH2 and ACTA2 of adult EPDCs cultured for 5 days in the presence of ActA (n 7) or ActA + SB (n 3), relative to CTRL. * p < 0.05, ** p < 0.01, ns not significant (mixed-effects analysis, Sidak's multiple comparisons test).
Source publication
The epicardium, the mesothelial layer covering the heart, is a crucial cell source for cardiac development and repair. It provides cells and biochemical signals to the heart to facilitate vascularization and myocardial growth. An essential element of epicardial behavior is epicardial epithelial to mesenchymal transition (epiMT), which is the initia...
Contexts in source publication
Context 1
... homo-or heterodimeric ligands are composed by combinations of two subunits encoded by INHBA and/or INHBB, which signal by binding to the type II receptors ACVR2A or ACR2B, and type I receptor ALK4. The presence of all components could be established in both adult and fetal EPDCs (Figure 2A, raw Ct values in italics indicate presence of mRNA transcripts). Furthermore, ALK4, INHBA and INHBB mRNA showed a trend towards higher expression in fetal EPDCs. ...
Context 2
... we validated the effect of Activin A signaling on epiMT markers in more detail. Fetal EPDCs treated with FST displayed a significant reduction of αSMA expression compared to control cells, confirming the prevention of epiMT (Figures 2B,C and Supplementary Figure S2 for DAPI). In adult EPDCs, the Activin A-induced phenotypical change towards a mesenchymal cell type was accompanied by a significant decrease of epicardial marker WT1, and an increase of mRNA expression of mesenchymal markers ACTA2 (encoding αSMA), POSTN and CDH2 confirming the occurrence of epiMT ( Figures 2D,E). ...
Context 3
... EPDCs treated with FST displayed a significant reduction of αSMA expression compared to control cells, confirming the prevention of epiMT (Figures 2B,C and Supplementary Figure S2 for DAPI). In adult EPDCs, the Activin A-induced phenotypical change towards a mesenchymal cell type was accompanied by a significant decrease of epicardial marker WT1, and an increase of mRNA expression of mesenchymal markers ACTA2 (encoding αSMA), POSTN and CDH2 confirming the occurrence of epiMT ( Figures 2D,E). However, this did not result in a large change in αSMA protein levels (Supplementary Figure S3). ...
