All images shown are of 3rd instar larval medulla. (A–A”) In situ hybridization of Ets65A mRNA (purple) in brains with D mutant clones marked by GFP (green). (B–B”) Immunostaining of Sox102F (red) and Oc (blue) in brains with D mutant clones marked by GFP (green). (C–C”) Immunostaining of Sox102F (red) in brains with slp mutant clones marked by GFP (green). (D) Immunostaining of Oc (blue) in brains with slp mutant clones marked by GFP (green). (E,E’) Immunostaining of Oc (blue) in brains with clones expressing eyRNAi driven by actin>Gal4 marked by GFP (green). (F) A model summarizing how temporal patterning generate different combinations of transcription factor expression in medulla neurons. Oc expression is started in late Ey stage, and is expressed in Notch-off progeny of Ey, Slp and D stages; Sox102F is expressed in Notch-on neurons born in the Slp and D temporal stages, and also in Notch-off neurons born in the D stage; Ets65A is expressed in Notch-off neurons born in the D stage, and continue to be expressed in progeny born at later stages. The Ap and Toy expression pattern are based on (Li et al., 2013).