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. 2020 Sep 1;21(17):6348. doi: 10.3390/ijms21176348

Figure 2.

Figure 2

Morphogen but not serum induces Ras/Raf/MEK/ERK cascade (MAPK)-dependent remodeling and dedifferentiation. (A) Myomesin and α-smooth muscle actin (SM-actin) fluorescence images after 6-day treatment. SM-actin serves as marker for dedifferentiation/fetal remodeling. Primary cultures of adult cardiomyocytes were untreated or treated with 20 nM isoproterenol (ISO), serum (Serum) and morphogens (Morpho). UO126 (5 µM) was added as an inhibitor of the MAPK pathway (+UO) 1h before experimental start. Note that knock-down of B-Raf (siBRaf) was performed for three days prior to stimulation. Single and double stars indicate p < 0.01 and p < 0.001, respectively. Black signs refer statistically to Con, blue signs to serum, and red compares Morpho (Morpho, Morpho siCon) statistically with and without UO/siB-Raf (+UO, siBRaf). A SM-actin positive fibroblast acts as positive control and is indicated by a white F. (B) Quantitative evaluation of the number of SM-actin positive cells and statistical analysis of (A). (C) Phase-contrast images of freshly isolated cells (Fresh), as well as Serum- (Serum 20 d) and Morpho-treated (Morpho 20 d) cardiomyocytes after 20 d showing long-term effects.