Fig. S4.

Pax6 knockdown and restoration after RNAi treatment in the time-course experiment. (A–D) Third-instar larval eye-antennal discs from tub-Gal80[ts]; DE-GAL4 > ey RNAi+ toy RNAi flies. Animals were kept at 18 °C AEL until the third-larval instar stage and were then transferred to 30 °C. Discs were dissected and immunostained to detect Ey and Toy proteins at 6, 8, 10, and 12 h ATS. Yellow arrows show the loss of Ey and orange arrows show loss of Toy in the dorsal compartment. (E–G) Third-instar larval eye-antennal discs from tub-Gal80[ts]; DE-GAL4 > toy RNAi + UAS-GFP flies. Animals were kept at 30 °C AEL until the third-larval instar stage, then being transferred to 18 °C. Discs were dissected and immunostained to detect Toy protein and GFP expression at 36, 40, and 44 h ATS. Red arrow indicates that Toy protein has not been restored at 36 h ATS. Green arrows indicate that at 40 h ATS, Toy expression has been restored in the dorsal half of the eye. Anterior is to the right. (Scale bars, 100 µm.)