DEV027318 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. HNT is not expressed in cone cells. Lateral posterior views of wild-type third instar larval eye discs angled to show an apical perspective. (A) This disc is stained with anti-ELAV (green) and anti-CUT (blue) antibodies to show the distinct R-cell precursor nuclei and more apical cone cell precursor nuclei. (B) In this disc, oriented as in A, the anti-ELAV antibody staining (green) in the R-cell nuclei overlaps with the anti-HNT antibody staining (red). There are no distinct red nuclei in the apical plane.

• Supplemental Figure S2 -

  Fig. S2. hnt mRNAi knockdown eye phenotypes. (A-A′′) Wild-type eye discs or (B-B′′) eye discs from pGMR>hnt RNAi (UAS-hnt mRNAi2A/pGMR-Gal4;hnt mRNAi2B/+) knockdown larvae stained with (A,B) anti-ROUGH (RO) antibody (red), (A′,B′) anti-HNT antibody (green) or (B′′) the R8 marker anti-SENS antibody (blue), which in our hands is a robust R2/5 marker and has much lower expression in R3/4. Even in the wild-type control (A), anti-ROUGH staining is often only found in two or three nuclei. In the knockdown mutant shown in B′′, HNT staining is retained in three cells including the SENS-positive R8 cell (blue/green overlap) and the two adjacent ROUGH-positive R2/5 cells (green/red overlap). The R3/4 nuclei do not retain HNT expression and are sometimes marked with ROUGH (red alone).

• Supplemental Figure S3 -

  Fig. S3. spa-Gal4 drives expression in a subset of the larval cone cell precursors. Apical views of a spa-Gal4/+; UAS-GFP/+ third instar larval eye disc stained with (A) anti-GFP antibody (green) alone, or (A′) together with anti-CUT antibody marking the cone cell precursors. The spa-Gal4 drives expression in a subset of the cone cell precursors in the larval eye disc.

• Supplemental Figure S4 -

  Fig. S4. Ectopic HNT is sufficient to induce CUT expression. Third instar eye discs stained with anti-CUT antibody. (A-C) Apical portion or (A′-C′) basal portion (excluding most of the CUT-positive, large basement glia nuclei found at the very bottom of the disc). (A,A′) Wild-type control disc showing (A) normal CUT expression in the cone cell precursors and (A′) almost no basal nuclei expressing CUT. (B,B′) Disc from a pGMR>hnt RNAi (UAS-hnt mRNAi2A/pGMR-Gal4;hnt mRNAi2B/+) larva with (B) fairly normal CUT expression in the cone cell precursors and (B′) ectopic CUT expression in many of the basal nuclei posterior to the furrow. (C,C′) Disc from a lz>hnt RNAi (lz-Gal4/Y; UAS-hnt mRNAi2A/+; UAS-hnt mRNAi2B/+) larva with (C) CUT expression in the cone cell precursors and (C′) ectopic CUT expression in some of the basal nuclei posterior to the furrow.