Fig. 5

Expression of nefma, nefmb, neff1 and inab in zebrafish evx1;evx2 double mutant and WT embryos. (A, B, D, E, G, H, J, K) Lateral views of (A, D, G, J) WT and (B, E, H, K) evx1^i232;i232;evx2^sa140;sa140 double mutant embryos (labeled evx1;evx2) at 30 h. Rostral, left. Dorsal, up. (C, F, I, L) Number of cells expressing (C) nefma, (F) nefmb, (I) neff1 and (L) inab in a precisely-defined spinal cord region adjacent to somites 6–10 at 30 h. Data are depicted as individual value plots and the n-values for each genotype are shown below. For each plot, the wider red horizontal bar depicts the mean number of cells, and the red vertical bar depicts the S.E.M. (mean numbers and S.E.M. values are listed in Table 1). All counts are an average of at least four embryos. Statistically significant comparisons are indicated with brackets and asterisks. *** P < 0.001. * P < 0.05. White circles indicate WT data and black circles indicate evx1;evx2 double mutant data. All data were analyzed for normality using the Shapiro–Wilk test. Data in C is not normally distributed and so a Wilcoxon-Mann–Whitney test was performed. Data sets in F, I and L are normally distributed and so the F-test for equal variances was performed, followed by a type 2 Student’s t-test (for equal variances). P-values are provided in Table 1. (C, I) There is a statistically significant reduction in the number of spinal interneurons expressing nefma and neff1, but not (F, L) nefmb and inab, in evx1;evx2 double mutant embryos. (A, B) nefma and (G, H) neff1 in situ hybridization experiments were performed with the molecular crowding reagent Dextran Sulfate. This was omitted for the (D, E) nefmb and (J, K) inab in situ hybridization experiments. Scale bar: 50 µm