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Fig. 6
a The hearts of AB fish were cryoinjured, and then the fish were maintained in untreated fish water (control) or in fish water containing 50 µM metformin or 100 µM CQ for 1?14 days. Sections were then prepared and stained with the MSB trichrome stain to show fibrin (in dark red) and collagen (in blue). V: ventricle; IA: injured area. Scale bar: 100 µm. b, c The hearts of Tg(cmv:GFP-LC3) fish were cryoinjured, and then the fish were maintained in untreated fish water (control) or in fish water containing 50 µM metformin or 100 µM CQ for 1-14 days. The hearts were then isolated, fixed, sectioned, and dual-immunostained with an anti-GFP antibody to label LC3 (in green) and an anti-vimentin antibody to label fibroblasts (in red), after which they were labeled with DAPI to show the nuclei (in blue). Scale bars: 50 µm (b). The numbers of GFP-LC3 puncta in the fibroblasts in the injured area of the control, metformin and CQ treated fish were quantified. The data are presented as mean ± SD, n = 3 hearts, *P < 0.05, **P < 0.01 vs control (c).