(A – E) Airyscan confocal optical sections of live 22 hpf transgenic jag1a:mScarlet zebrafish injected with emilin3a:GFP (A and B), emilin3a:GFP-p2a-her6 (C and D) or emilin3a:GFP-p2a-her9 (E and F). DNA constructs were injected at the one-cell stage together with I-SceI protein. (B, D, F) show the boundary of GFP segmentation in A, C, and E, respectively, and manual outline of the notochord. (G), Quantification of jag1a:mScarlet intensity inside GFP-positive cells segmented as exemplified in (B, D, F). Values in the plot represent the intensity of jag1a:mScarlet inside segmented cells divided by the jag1a:mScarlet intensity inside the notochord outside of the segmented cells. Each point represents an individual fish (n = 12 GFP, n = 7 GFP-p2a-her6, n = 11 GFP-p2a-her9). Two-tailed p-values are shown in the plot. (H), Airyscan confocal sections of embryo at 22 hpf injected with Cas9 together with a control guide (H) or Cas9 together with her6/her9 gRNAs (I). (J) Quantification of the average number of jag1a-positive cells directly adjacent to each jag1a-positive cell. For each individual fish, we count how many jag1a-positive cells are adjacent to each jag1a-postive cell, and then calculate the average for that fish. This value would be equal to 2 in case all the cells are jag1a-positive, and zero if no jag1a-positive cell is adjacent to another jag1a-positive cell. Each individual point in the plot represents the average value for an independent fish (n = 7 control, n = 8 her6/her9 KOs). Two-tailed p-value is shown in the plot. Scale bars, 20 μm.