Fig. 2

NAD+ and EmergenC supplementation at gastrulation do not significantly improve the UPR or vascularization in fkrp morphants. (A–C1) Anterior left, dorsal top, side-mounted 3 dpf embryos expressing Tg (ef1α:xbp1δ-GFP). Fluorescence intensity was kept constant within an experiment (see methods). Numbered panels are merged with the brightfield channel. (A, A1) Control embryo. Note the low relative expression of Xbp1 compared to morphants. (B, B1) fkrp morphant. (C, C1) fkrp morphant treated with EmergenC at gastrulation. Note that fluorescence intensity is similar to that of untreated morphant. (D) Quantification of Xbp1 fluorescence intensity as a percent of the wild-type value for all groups imaged. Fluorescence intensity is significantly increased in morphants. There is no significant difference in fluorescence intensity between untreated morphants (n = 15 embryos) and morphants receiving EmergenC (n = 21 embryos). (E–G) Anterior left, dorsal top, side-mounted 2 dpf embryos expressing Tg (fli1:EGFP) focused on the ISVs. (E) Control embryo. (F) fkrp morphant embryo. Note that some ISVs are truncated (white arrowhead). (G) fkrp morphant embryo treated with NAD+ at gastrulation. Truncated ISVs are still present in NAD+ treated morphants (white arrowhead). (H) Quantification of ISV length. ISV length is reduced in fkrp morphants (n = 201 vessels) compared with uninjected controls (n = 84 vessels). NAD+ supplementation (n = 166 vessels) does not rescue ISV length in fkrp morphants. Scalebars are 50 μm. *p < 0.05, **p < 0.01, ***p < 0.001, ns non-significant