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. 2020 Jul 24;21(15):5258. doi: 10.3390/ijms21155258

Figure 1.

Figure 1

Hepatocyte growth factor (HGF) protects cardiomyoblasts against doxorubicin (Doxo)-induced DNA damage and apoptosis and the inhibition of Erk1,2 (extracellular signal-regulated kinase 1,2) dampens this cardioprotection. (a) Scheme of the experimental protocol used for treatments. H9c2 cardiomyoblasts were incubated with Doxo (25 μM) for 1 h (Doxo pulse) or pretreated for 4 h with HGF (0.5 nM) alone or HGF+PD (PD98059, Erk inhibitor) and exposed to Doxo in the last hour. Then, the cells were replaced with fresh low serum medium (recovery time). (b) Control, Doxo- and Doxo+HGF-treated cells were analyzed at different time points of the recovery time (0, 2, 6 12 and 24 h). (c) Control, Doxo-, Doxo+HGF- and Doxo+HGF+PD-treated cells were analyzed at 2 and 24 h of recovery time. P-Erk1,2 and total Erk2 proteins were detected by Western blotting. αtubulin was used as the loading control in all Western blots. (d) The level of γH2AX protein was analyzed by flow cytometry. (e) The level of cleaved caspase 3 protein was analyzed by flow cytometry. Data are representative results of three independent experimental replicates.