ZFIN Image: Sorrells et al., 2018, Fig. 4

Image

Figure Caption

Fig. 4

RNASEH1 reverses R-loops, DNA DSBs and apoptosis in spliceosomal mutant zebrafish. (A) Schematic of the RNASEH1 overexpression experiment. (B) Quantification of R-loop levels in cells isolated from 24 hpf Tg(hsp70:M27RNASEH1-GFP)-negative and Tg(hsp70:M27RNASEH1-GFP)-positive sf3b1^hi3394aTg mutant and wild-type siblings following heat shock induction. (C) Confocal images showing immunofluorescence of nuclei (DAPI) and DNA damage (γH2AX) in cells isolated from 24 hpf Tg(hsp70:M27RNASEH1-GFP)-negative and Tg(hsp70:M27RNASEH1-GFP)-positive sf3b1^hi3394aTg mutant and wild-type siblings. The lower panel is a merged image with an inset of a single cell to better show γH2AX staining. (D) Quantification of γH2AX fluorescence intensity in sf3b1^hi3394aTg mutant and wild-type siblings. (E) Representative flow cytometry plots showing active caspase-3 levels on the y-axis and forward scatter (FSC) on the x-axis. (F) Quantification of active caspase-3 levels in 24 hpf Tg(hsp70:M27RNASEH1-GFP)-negative and Tg(hsp70:M27RNASEH1-GFP)-positive sf3b1^hi3394aTg mutant and wild-type siblings following heat shock induction. For images, 63× magnification with 6.5× zoom. Scale bars: 5 μm. Error bars represent s.e.m. P-values were calculated by one-way ANOVA with Sidak's multi-testing correction. *P<0.05; **P<0.01; ***P<0.001; **** P<0.0001; n.s., not significant. For D and F, data represent normalized values across three replicates.

Figure Data

Expression / Labeling details

Phenotype details

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Dis. Model. Mech.