Fig. 7.

NF-κB regulates Notch targets downstream of Crip genes. (A) Experimental design schematic depicts the addition of DMSO and CAPE to wild-type and crip^DM embryos at 24 hpf with fixation at 28 hpf for confocal microscopy or at 32 hpf for cmyb in situ hybridization (ISH). (B-E) ISH for cmyb at 32 hpf illuminates reemergence of HSPCs in CAPE-exposed (n=15) (E) compared to DMSO-exposed (n=18) (D)  crip^DM embryos. Images from DMSO- (n=16) (B) and CAPE-exposed (n=19) (C) wild-type embryos are also depicted. Lateral views, anterior to the left. Scale bar: 100 µm. (F) Quantification of cmyb signals in the dorsal aorta (DA) from B-E using pixel intensity analysis shows a statistically significant boost in the CAPE-exposed compared to the DMSO-exposed crip^DM embryos. Unpaired nonparametric Mann–Whitney U-test yields ****P<0.0001 between DMSO- and CAPE-exposed wild-type embryos, between DMSO-exposed wild-type and crip^DM embryos, and between DMSO- and CAPE-exposed crip^DM embryos. (G-K) Confocal images of DMSO-treated wild-type (n=18) (H) and crip^DM (n=22) (J) and CAPE-treated wild-type (n=18) (I) and crip^DM (n=32) (K) embryos carrying Tg(kdrl:mCherry);Tg(tp1:EGFP) at 28 hpf underscore a rescue of kdrl:mCherry⁺tp1:EGFP⁺ cell number in CAPE-exposed crip^DM embryos. Lateral views, anterior to the left. Arrows indicate kdrl:mCherry⁺tp1:EGFP⁺ double positive cells in the floor of the dorsal aorta. Scale bar: 25 µm. Quantification of kdrl:mCherry⁺tp1:EGFP⁺ cells in H-K (G) shows a statistically significant decrease in the CAPE-exposed compared to the DMSO-exposed crip^DM embryos. Unpaired nonparametric Mann–Whitney U-test yields ****P<0.0001 between DMSO-exposed wild-type and crip^DM embryos and between DMSO- and CAPE-exposed crip^DM embryos. Mean and standard error of each dataset are shown. ns, not significant.