Figure 3.

Met agonist mAbs and HGF protect H9c2 cardiomyoblasts from CoCl₂-induced apoptosis. Cells were untreated (NT, white) or treated with 300 μM CoCl₂ (black), CoCl₂+DN30 (light grey), CoCl₂+DO24 (middle grey) and CoCl₂+HGF (dark grey). MAbs (100 nM) and HGF (0.5 nM) were concomitantly added to CoCl₂ for 24 h (a) or for 48 h (b–d). Cell viability (a) and cell count (b) were measured. In a, the separated columns on the right represent cells treated with CoCl₂+each Met agonist in the presence of PHA Met inhibitor (500 nM). Results are expressed as the percentage of MTT or cell number reduction, relative to NT control. (c) The number of DAPI-stained pyknotic nuclei on the total amount of nuclei was counted. For all treatments, the mean±S.D. was obtained from 10 independent experiments. (d) Caspase-3 protein cleavage was analysed by WB. The values show the ratio between the densitometric quantification of cleaved and total caspase-3 bands. Values are expressed as fold relative to NT control and representative images are shown below the graph. The mean±S.D. was calculated in three independent experiments, except for the DAPI stain (c). For t-test each group of samples was compared with the CoCl₂-treated cells. *P<0.05, **P<0.01 and ***P<0.005