However, the remaining horizontal cells expressed Sall3, Prox1 and NF165 (see Fig. S2M,P in the supplementary material, arrows).
In P0-14DIV Sall3−/− explants, a few Arr3-positive cells were seen, although many fewer than in wild-type explants (see Fig. S5I,L in the supplementary material, arrow).
This drastic reduction in Sop expression was also observed in 14 day (P0-14DIV) Sall3−/− retinal explants (see Fig. S5F in the supplementary material, arrow).
Indeed, CARMIL and Myosin 1E were co-localized in the spots, as well as at the tip of lamellipodia (Fig. 2A,E arrows; supplementary material Fig. S4C dark blue circle; supplementary material Movie 5).
When we expressed the Rho1 reporter with RhoGEF2 in the posterior compartment of eye discs under the control of GMR-GAL4, RhoGEF2 was localised apically and greater accumulation of GFP was observed in the apical-lateral region of these cells (arrows, supplementary material Fig. S2Bv,Bviii) compared with the control (arrows, supplementary material Fig. S2Aiv,Avi; quantified in S2C).
In contrast, knockdown of Myosin II did not alter the Raf GOF phenotype; ectopic differentiation was still observed anterior to the MF (arrows, supplementary material Fig. S8C-Cii) and some photoreceptor nuclei were basally localised (arrows, supplementary material Fig. S8D-Dii), but F-actin levels were not affected compared with the surrounding wild-type tissue (supplementary material Fig. S8E).
In addition to the described effect on vascular and lymphatic development, tmem33 morphants exhibited increased glomerular size (Supplementary Fig. 7m o, arrows) and homozygous tmem33sh443 mutants were protected against the effect of the tmem33 morpholino on glomerular size (Supplementary Fig. 7m o, arrows).
The superimposition (FAK/Actin; yellow) of FAK (green) and actin (red) show FAK associated with actin filaments at the leading edge of the protrusion front during the 60-min time course (arrows) (Supplementary Movie 16).
We also observed that this most anteriorly located Cerl-GFP-expressing cell extended projections towards the anterior region and initiated migration in that direction (Fig. 6a, arrows, Supplementary Movie 7).
TIAF1 debris, rather than de novo synthesized materials, is shown (see white arrows; Supplementary Figure S12).
Co-localization with NF-H further showed that caspase 3 was affecting neuronal axons (Supplementary Figure S1, arrows).
