miR26a knockdown embryos have increased endothelial pSmad1.A-D) Ventral view confocal projections of the 4 dpf ventral aorta (dotted outline). Endothelial nuclei (fli1a:nEGFP; A-D, arrowheads) and pSmad1/5/9 (pSmad1,white A’- D’) and overlay (magenta, A”- D”) in 4 dpf Scr. Control (A), miR26a MO (B), dCas9 control (C) and miR26a CRISPRi (D) embryos. Yellow arrowheads indicate double positive pSmad1 + fli1a:nEGFP nuclei in the ventral aorta. E) Quantification of total number of fli1a:nEGFP nuclei in the ventral aorta. F-G) Quantification of the percentage of double pSmad1; fli1a:nEGFP positive nuclei in miR26a morphants (F) and miR26a CRISPRi (G) embryos. N = 3 experiments. Total embryos are as follows: Scr. Control MO n = 14, miR26a MO n = 14, dCas9 control n = 9 and miR26a CRISPRi n = 10. Student's two-tailed t-test, p***< 0.0001 and p ****< 0.00001 as compared to WT, error bars = SD. Scale bar: represents 50μm.
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