Overexpression of mdka partially rescues the liver regeneration defect in the pdgfra mutant. (A) Whole-mount in situ hybridization images showing the expression of mdka in wild-type and mild pdgfra mutant livers at 8 h after injury (at regeneration 8 h; R8 h) and R24 h. The white dashed line outlines the liver region. (B) Whole-mount in situ hybridization images showing the expression of mdka in dominant-negative pdgfra− and dominant-negative pdgfra+ livers at R8 h and R24 h. The liver region is outlined. (C) Fluorescent in situ hybridization and antibody staining showing mdka expression in the GFP+ HSCs in wild type and mild pdgfra mutant at R8 h and R24 h under the Tg(lfabp:DenNTR; hand2:GFP) background. Scale bars: 50 μm. (D) Experimental scheme illustrating the treatment of Mtz or DMSO and the mdka overexpression by heat shock (HS; abbreviated as mdka+) under the Tg(lfabp:DenNTR; hsp70L:mdka-HA-p2a-Venus) background. (E) Confocal projection images showing the liver regeneration at R48 h. Scale bars: 50 μm. (F) Statistical analysis of intensity of Dendra2. n=12 (wild type), n=9 (mild pdgfra−/− and mild pdgfra−/−; mdka+) and n=11 (wild type; mdka+). (G) Whole-mount in situ hybridization images showing the expression of gc and cp at R48 h. (H,I) Statistical analysis of intensity of gc and cp expression. n=12 (gc wild type), n=8 (gc mild pdgfra−/−) and n=11 (gc mild pdgfra−/−; mdka+), n=12 (cp wild type) n=8 (cp mild pdgfra−/−) and n=7 (cp mild pdgfra−/−; mdka+). (J) Single-optical section images showing Bhmt and Dendra2 expression at R48 h. Scale bars: 50 μm. (K) Statistical analysis of percentage of Bhmt+ cells among Dendra2+ cells. n=9 (wild type and mild pdgfra−/−), and n=8 (mild pdgfra−/; mdka+). Numbers indicate the proportion of larvae exhibiting the expression shown. ns, no significant difference; *P<0.05, ***P<0.001, ****P<0.0001 (unpaired t-test). Data are mean±s.e.m.
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