DEV010876 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Additional phenotypes of babo mutants. An eye-antenna disc of a yw (A) late third instar larva stained by 24B10 antibody (red), Elav antibody (magenta), and phosphorylated histone H3 (p-H3) antibody (green) compared with an eye-antennal disc of a babo^32/52 mutant (B). The size of the eye-antenna disc of the babo^32/52 mutant is not much smaller, nor does it have a reduced number of M-phase cells compared with the yw control. Note, however, that there are ectopic p-H3 staining cells in the morphogenetic furrow of the babo^32/52 mutant (arrows). (C) In the weakest babo^32/52 larva, lamina cartridge neurons (Dac⁺, green) are present in approximately normal numbers in some columns (arrow), but the total size of the lamina is reduced. (D) Expression of Drosophila actβ (dAct) mRNA in a wild-type yw wondering third instar larva brain lobe (arrows) and eye disc (arrowhead) as revealed by in situ hybridization. (E) dpp mRNA expression in yw third instar larva brain lobes. Expression is seen in limited groups of neuroepithelial cells in both outer (arrowheads) and inner (arrows) proliferation centers. (F) dpp mRNA expression is unaltered in the brain lobes of a babo^32/52 mutant.

• Supplemental Figure S2 -

  Fig S2. The number of central brain neuroblasts is reduced in babo mutants. (A) Bar chart of average number of Dpn⁺ central brain neuroblasts and average number of proliferating neuroblasts (Dpn⁺, phospho-histone H3⁺). Wild-type numbers in blue bars; babo^32/52 numbers in red bars. Wild-type white prepupa (wpp) central brain has an average of 99.87 large Dpn⁺ neuroblasts and an average of 26.9 proliferating neuroblasts. babo^32/52 wpp central brain has an average of 83 large Dpn⁺ neuroblasts (17% fewer) and an average of 23.5 proliferating neuroblasts. P value using Student’s t-test. (B) Wild-type wpp brain lobe. The large optic lobe is outlined (white dots). Anterior is up; lateral is right; medial midline is left. Scribble (Scrib, blue) outlines all cell cortices in the wpp optic lobe and central brain. Elav (red) marks all maturing neurons in both central brain and optic lobe. Prospero (Pros, green) is detected in ganglion mother cells and maturing neurons of both central brain and optic lobe. Differentiating neurons in the optic center are seen (arrowheads). Early GMCs are positive for Pros in central brain lobe (arrow). (C) A babo^32/52 mutant wpp stained with the same antibodies as in B. Note that very few Elav⁺ neurons are seen in the brain lobe in the babo^32/52 mutant. In addition, the Pros⁺ GMCs in the inner proliferation center are not present.

• Supplemental Figure S3 -

  Fig. S3. Characterization of the WorGal4 expression in wondering third instar larval brains. (A-D) Identical sections of an optic lobe showing WorGal4-GFP (A), Miranda (B), Elav (C) and a merged view (D). Arrowheads in D indicate expression of Wor in subsets of neuroblasts within the OPC and IPC. (E-H) Enlargements of the boxed area of D, illustrating expression of Wor in a single central brain neuroblast and several of its GMC daughters.