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( A, B ) Immunostaining of cyclobutene pyrimidine dimer (CPD) shows a similar DNA damage level induced by UV in surface fish and Pachón cavefish embryonic fibroblasts. White circles indicate the nuclear area by DAPI staining. Orange indicates CPD. Scale bar, 40 μm. P -values were determined by two-way ANOVA: F =0.09703, p =0.7586. ns p =0.6404, **** p <0.0001. ( C, D ) Western blot <t>of</t> <t>γH2AX</t> indicates a diminished DNA damage response in Pachón cavefish embryonic fibroblasts compared to surface fish cells. ( E, F ) Flow cytometry images and quantification for host cell reactivation assays in surface fish and Pachón cavefish embryonic fibroblasts. Red line sets the green fluorescent protein <t>(GFP)</t> positive signal threshold. P -values were determined by unpaired t -test. ( G ) Representative GFP images for host cell reactivation assays in surface fish and Pachón cavefish embryonic fibroblasts. Scale bars, 500 μm. Figure 4—source data 1. Original gels for data presented in . Figure 4—source data 2. Original western blots for data presented in .
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( A, B ) Immunostaining of cyclobutene pyrimidine dimer (CPD) shows a similar DNA damage level induced by UV in surface fish and Pachón cavefish embryonic fibroblasts. White circles indicate the nuclear area by DAPI staining. Orange indicates CPD. Scale bar, 40 μm. P -values were determined by two-way ANOVA: F =0.09703, p =0.7586. ns p =0.6404, **** p <0.0001. ( C, D ) Western blot of γH2AX indicates a diminished DNA damage response in Pachón cavefish embryonic fibroblasts compared to surface fish cells. ( E, F ) Flow cytometry images and quantification for host cell reactivation assays in surface fish and Pachón cavefish embryonic fibroblasts. Red line sets the green fluorescent protein (GFP) positive signal threshold. P -values were determined by unpaired t -test. ( G ) Representative GFP images for host cell reactivation assays in surface fish and Pachón cavefish embryonic fibroblasts. Scale bars, 500 μm. Figure 4—source data 1. Original gels for data presented in . Figure 4—source data 2. Original western blots for data presented in .

Journal: eLife

Article Title: Elevated DNA damage without signs of aging in the short-sleeping Mexican cavefish

doi: 10.7554/eLife.99191

Figure Lengend Snippet: ( A, B ) Immunostaining of cyclobutene pyrimidine dimer (CPD) shows a similar DNA damage level induced by UV in surface fish and Pachón cavefish embryonic fibroblasts. White circles indicate the nuclear area by DAPI staining. Orange indicates CPD. Scale bar, 40 μm. P -values were determined by two-way ANOVA: F =0.09703, p =0.7586. ns p =0.6404, **** p <0.0001. ( C, D ) Western blot of γH2AX indicates a diminished DNA damage response in Pachón cavefish embryonic fibroblasts compared to surface fish cells. ( E, F ) Flow cytometry images and quantification for host cell reactivation assays in surface fish and Pachón cavefish embryonic fibroblasts. Red line sets the green fluorescent protein (GFP) positive signal threshold. P -values were determined by unpaired t -test. ( G ) Representative GFP images for host cell reactivation assays in surface fish and Pachón cavefish embryonic fibroblasts. Scale bars, 500 μm. Figure 4—source data 1. Original gels for data presented in . Figure 4—source data 2. Original western blots for data presented in .

Article Snippet: Consistent with the γH2AX findings, the UV-damaged plasmid-transfected cavefish cells displayed a substantially lower GFP signal recovery (~22% relative to control) than the surface fish cells (~49% relative to control) ( ).

Techniques: Immunostaining, Staining, Western Blot, Flow Cytometry, Host-Cell Reactivation