Figure 3
DAC is required for CLK cross-reactivity in non-oscillator cells. CNSs were dissected from dac03 mutant L3 larvae collected at ZT21 and ZT9, immunostained with DAC, CLK GP47, and PER antibodies, and imaged by confocal microscopy. (A-D, E-H) Projected Z-series images of the CNS from L3 larvae are shown, where dorsal is at the top. (A-D') CNS from an L3 larva collected at ZT21 was imaged for DAC (A), CLK (B), PER (C) or DAC, CLK and PER (D) immunostaining. Boxed region in panels A-D is magnified in panels A'-D', respectively. (B'-C') Brackets denote CLK and/or PER IR in DN1* neurons, and arrowheads denote CLK and/or PER IR in s-LNv and DN2 neurons. DN1*, s-LNv, and DN2 neurons are as defined in the legend for Fig. 2. Co-localization of CLK (red) and PER (blue) appears as white. (E-H') CNS from an L3 larva collected at ZT9 was imaged for DAC (E), CLK (F), PER (G) or DAC, CLK, and PER (H) immunostaining. Boxed region in panels E-H is magnified in panels E'-H', respectively. (F'-H') Arrowheads denote CLK and/or PER IR in DN1*, s-LNv, and DN2 neurons. Images are representative of three independent experiments at each time point. At least three larval CNSs were examined for each experiment. Z-series images are projections over 33 optical sections at 2.5 μm per optical section.