Fig. 5

mrpl40 mutants have fewer NSPCs and a reduction in NSPC proliferation in the dorsal proliferative zone.

A–D PCNA and DAPI antibody staining in wildtypes (A), mrpl40 mutants (B), and prodha mutants (C) in dorsal proliferative zone. mrpl40 mutants have fewer PCNA+ cells than wildtypes. prodha mutants have similar numbers to wildtypes. Quantification in D. One-way ANOVA, p-values are displayed on the plots. Siblings (n = 21), mrpl40 mutants (n = 10), prodha mutants (n = 13) form two biological replicates. Both mrpl40 and prodha mutants appear to have increased ventricle size (yellow arrows). Images are single confocal slices. Scale bars 15 μm. E PH3 staining in dorsal proliferative zone in siblings, mrpl40 mutants, and prodha mutants.. mrpl40 mutants show a decrease in the number of Glast−/PH3+ cells compared to siblings. Unpaired two-way Student’s t test, p-values displayed on the plots. mrpl40 mutants (n = 19) and siblings (n = 25). prodha mutants (n = 25) and siblings (n = 26) from four biological replicates. F–H Sox2/PCNA antibody staining in wildtypes (E) and mrpl40 mutants (F) in dorsal proliferative zone. mrpl40 mutants show a reduction in the number of Sox2−/PCNA+ cells but no difference in the number of Sox2+/PCNA+ cells (G). Quantification in G. Unpaired two-way Student’s t test, p-values displayed on the plot. mrpl40 mutants (n = 10) and siblings (n = 12) from three biological replicates. Images are single confocal slices. Dotted line indicates boundary of Sox2+ cells. Scale bars 15 μm. I–J Proliferative cells in dorsal hindbrain of mrpl40 mutants (I) display elongated nuclei compared to wildtypes (H). Images are single confocal slices. Scale bars 10 μm. K–M PH3/PCNA antibody staining in wildtypes (J) and mrpl40 mutants (K) in dorsal proliferative zone. mrpl40 mutants show a reduction in the number of PH3+ cells located both centrally along ventricle and more peripherally. Quantification in L. Unpaired two-way Student’s t tests, p-values displayed on the plot. mrpl40 mutants (n = 14) and siblings (n = 15) from three biological replicates. Images are maximum projections from confocal stacks. Scale bars 20 μm.