pdgfra inactivation limits HSC expansion during liver regeneration. (A) Experimental scheme illustrating the treatment of Mtz or DMSO and analysis at 6 dpf, 8 h after injury (at regeneration 8 h; R8 h), R24 h and R48 h under the Tg(lfabp:DenNTR; hand2:GFP) background. (B) Confocal projection images showing the antibody staining of GFP in wild type and mild pdgfra mutant at 6 dpf, R8 h, R24 h and R48 h. Scale bars: 50 μm. (C) Statistical analysis of the number of HSCs. 6 dpf: n=8 (wild type) and n=5 (mild pdgfra−/−); R8 h: n=10 (wild type) and n=14 (mild pdgfra−/−); R24 h: n=8 (wild type) and n=9 (mild pdgfra−/−); R48 h: n=9 (wild type) and n=8 (mild pdgfra−/−). (D) Single-optical section images showing EdU and GFP antibody staining at R8 h in wild type and mild pdgfra mutant. Arrows indicate EdU− HSCs; arrowheads indicate EdU+ HSCs. Scale bars: 50 μm. (E) Statistical analysis of the percentage of EdU+ cells among GFP+ cells. n=9 (wild type) and n=8 (mild pdgfra−/−). (F) Experimental scheme illustrating treatment with Mtz or DMSO, and heat shock and analysis at R0 h and R24 h. (G) Confocal projection images showing the antibody staining of GFP in dominant-negative pdgfra− and dominant-negative pdgfra+ at R24 h. Scale bars: 50 μm. (H) Statistical analysis of number of HSCs. n=6. (I) Single-optical section images showing PCNA and GFP antibody staining at R0 h in dominant-negative pdgfra− and dominant-negative pdgfra+. Arrows indicate PCNA− HSCs; arrowheads indicate PCNA+ HSCs. Scale bars: 50 μm. (J) Statistical analysis of the percentage of PCNA+ cells among GFP+ cells. n=8. 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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