Fig. 3.

CLK expression in DOL cells promotes GFP-CYC accumulation. The 3.0cry-Gal4, GFP-cyc (3.0cry-G4, GFP-cyc), and 3.0cry-Gal4, GFP-cyc; UAS-Clk/+ (3.0cry-G4, GFP-cyc; U-Clk) flies were entrained in LD for at least 3 d and collected at ZT2. Immunostaining with GFP and PER antibodies was performed on dissected brains and imaged by confocal microscopy. Projected Z-series images of right brain hemispheres are shown, where lateral is right and dorsal is top. Pacemaker neuron groups are as defined in the text, and DOL cells are as defined in Fig. S1. Colocalization of GFP (green) and PER (red) is shown as yellow. (A–C) A 76-μm projected Z-series image of a 3.0cry-G4, GFP-cyc fly brain immunostained with GFP (A), PER (B), or GFP and PER (C). (D–F) An 86-μm projected Z-series image of a 3.0cry-G4, GFP-cyc; UAS-Clk fly brain immunostained with GFP (D), PER (E), or GFP and PER (F). GFP and PER immunostaining is detected in the indicated groups of pacemaker neurons and DOL cells, as well as in additional ectopic locations. (Scale bar, 10 μm.) All images are representative of six or more brains.