Fig. 3 - Supplemental 1

Analysis of proliferation and apoptosis in the OPs of sly mutants and control siblings. (A-D’) Immunostaining for Phospho-histone H3 (magenta) to label dividing cells on Tg(neurog1:GFP) embryos (green) at 16 hpf and 21 hpf (A-B’ show images at 21 hpf, dorsal view), and on Tg(omp:meYFP) embryos (green) at 28 and 36 hpf (C-D’ show images at 36 hpf, frontal view). In A-B’ the OP is surrounded by a dotted line. Arrows = instances of Phospho-histone H3 positive cells in the placodal Tg(neurog1:GFP)+ or Tg(omp:meYFP)+ clusters. (E–H) Number of Phospho-histone H3 positive cells in the OP at 16 hpf (n=19 controls; n=8 mutants), 21 hpf (n=21 controls; n=6 mutants), 28 hpf (n=8 controls; n=7 mutants) and 36 hpf (n=9 controls; n=9 mutants) in control embryos and sly mutants. Unpaired, two-tailed t test. (I-L’) Immunostaining for Activated Caspase 3 (magenta) to label apoptotic cells/debris on Tg(neurog1:GFP) embryos (green) at 16 hpf and 21 hpf (I-J’ show images at 21 hpf, dorsal view), and on Tg(omp:meYFP) embryos (green) at 28 and 36 hpf (K-L’ show images at 36 hpf, frontal view). In I-J’ the OP is surrounded by a dotted line. Arrows = instances of Activated Caspase 3 positive cells or debris in the OP or in contact with the placodal Tg(neurog1:GFP)+ or Tg(omp:meYFP)+ clusters. (M–P) % of left OPs with zero, one or two and more Activated Caspase 3 positive cells or debris in the OP or very close to the OP at 16 hpf (n=25 controls; n=9 mutants), 21 hpf (n=25 controls; n=9 mutants), 28 hpf (n=8 controls; n=10 mutants) and 36 hpf (n=10 controls; n=10 mutants) in control embryos and sly mutants. Chi2 test. (Q, R) Total number of YFP+ cells in control and mutant Tg(omp:meYFP)+ OPs at 28 hpf (Q) and 36 hpf (R) (n=3 control placodes; n=3 mutant placodes for each stage). Unpaired, two-tailed t test.