|
Figure 4 (A, B) Lateral confocal projections of Tg(6xHsa.NFKB:EGFP)nc1 embryos reporting NfkB signalling levels at 48hpf for WT (A) and hai1afr26 (B). (C) qPCR of cDNA levels of NfkB target gene nfkbiaa in hai1afr26 vs. sibs at 48hpf. n = 3, 200 embryos pooled in each, t-test *p<0.05. (D–E′) Projected confocal images of the tail fins of 48hpf Tg(mpx:eGFP)i114; hai1ahi2217 embryos, immunostained for TP63 (magenta) and eGFP (green) (D, E) with corresponding DIC image (D′, E′). Embryos were either mutant for ikbkg (ikbkgsq304, E–E′) or heterozygous (ikbkg+/sq304; D–D′). (F) Counts of eGFP-positive neutrophils in the fins at 48hpf of hai1ahi2217; ikbkg+/sq304 and hai1ahi2217; ikbkgsq304. Embryos were hemizygous for Tg(mpx:eGFP)i114. n = 9; t-test; ***p<0.001. (G–I) Lateral DIC images of the trunk of hai1a+/hi2217; ikbkg+/sq304 (G), hai1ahi2217; ikbkg+/sq304 (H), and hai1ahi2217; ikbkgsq304 (I). Loss of IKBKG does not rescue epidermal defects of hai1a mutants (arrowheads). (J, K) Lateral confocal projections of Tg(6xHsa.NFKB:EGFP)nc1 embryos reporting NfkB signalling levels at 32hpf of hai1ahi2217 uninjected (J) or injected with duox MO (K). Loss of H2O2 reduces NfkB signalling levels in hai1a mutants. Scale bars: (A, K) = 200 µm; (D′, I) = 50 µm.