The number of proliferating cardiomyocytes and the accumulation of fibroblasts are reduced (but apoptosis is not affected) in GM6001-treated fish following cryoinjury. (A) Representative paraffin sections showing the margin between the intact ventricle and injured area (IA) in (Aa) DMSO control and (Ab) GM6001-treated fish, which were immunolabelled with MF20 and an anti-PCNA antibody to label the cardiomyocytes and proliferating cells, respectively. The sections were also co-labeled with DAPI to label the nuclei. (Ac) Bar chart to show the numbers of proliferating cardiomyocytes in the margin between the intact ventricle and IA in control and GM6001-treated fish. (Ba,Bb) Representative paraffin sections were immunolabelled with an anti-vimentin antibody to label fibroblasts, after which (Bc) the percentage of vimentin expression in the scar of the DMSO- and GM6001-treated fish at 4 dpc was determined. (C) Representative paraffin sections of intact and cryoinjured hearts after the fish were injected with either: (Ca,Cb) DMSO (controls) or (Cc,Cd) GM6001 (prepared in DMSO). Cells undergoing apoptosis were stained with TUNEL (pink). (Ce) Bar chart to show the percentage of apoptotic cells in the intact ventricle, and in the injured area at 1 dpc to 7 dpc. In (Ac and Bc), the data are expressed as the mean ± standard deviation of n = 4–6 hearts, and the asterisks indicate GM6001 data that are significantly different from the DMSO controls at p < 0.05 (*) and p < 0.01 (**), two-tailed t-test. In (Ce), the data are expressed as the mean ± standard deviation of n = 4 hearts, and n.s. indicates data that are not statistically different, one-way ANOVA with LSD Post-hoc test.
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