Fig. 7: GATA2 expression and IEE birth epoch are expanded in developing cRE1^dup/+ hindbrain.

a–h, E14.5 WT (a–d) and cRE1^dup/+ (e–h) hindbrain sections at r4 (a,b,e,f) and r6 (c,d,g,h) axial levels showing immunofluorescence with ISL1 (blue) and GATA3 (green) (a–h) together with GATA2 (red: a,c,e,g) or NR2F1 (red: b,d,f,h). Ectopic dorsal r4MNs are present in e and f compared with a and b. Dotted yellow and blue rectangles (a,b,e,f) surround IEE VEN and OCN regions, respectively, and are magnified below. Dashed white squares (c,d,g,h) surround facial nuclei and are magnified below. White arrows show OCNs and white arrowheads FBMNs. The borders of the hindbrain are outlined in gray. Scale bars, 200 μm (a,c) apply to a, b, e and f, and c, d, g and h, respectively (n = 3 (a,c), 3 (b,d), 8 (e,g) and 7 (f,h) embryos). i–k, Schematics of E14.5 hindbrain cytoarchitecture based on a–h as viewed ventrally (i) and in cross-section at the level of r4 (j) and r6 (k) in WT (left side) and cRE1^dup/+ (right side) of hindbrains. l–o, Quantification of E14.5 r4MN transcriptional and positional identity in cRE1^dup/+ and WT littermates detected in confocal z stacks. Unilateral counts are presented; each point represents an individual embryo and each color a litter (color coded A–F) (n = 9 cRE1^dup/+ and nine WT littermate pairs from six litters). On average per side, WT versus cRE1^dup/+ embryos had: 9,470 versus 10,422 r4-born MNs (l); 903 versus 4,405 IEEs (m); 8,408 versus 5,691 FBMNs (m); 719 versus 2,478 OCNs (n); 184 versus 1,927 VENs (n); and 7,721 versus 2,098 FBMNs completing migration into ventral r6 (o). In the box plot, the center line is the median, the box limits represent 50% of the values and the whiskers represent 98% of the values. p, The r4MN birthdating in the 18-mouse cohort in l–o using in utero labeling of mitotic cells with thymidine homolog EdU in IEE (OCN + VEN) and FBMN (FBMN + r4 ectopic); definitions as per m. Point is the mean. For l–p, all indicated P values are calculated using two-sided, pairwise Student’s t-test without correcting for multiple testing; the error bar = ± s.e.m. See also Extended Data Figs. 8 and 9.