Immunostaining of Drosophila Pupal Eye

Immunostaining of the pupal eye was carried out as described [14] (link). Rat anti-Kirre (1∶5000) and rabbit anti-Hbs AS14 (1∶2500) were used as previously described [23] (link). Other primary antibodies: mouse anti-Rst Mab24A5.1(1∶100) [33] (link), rabbit anti-Sns (1∶300) [34] (link) and rabbit anti-lacZ (1∶2000; 5 Prime→3 Prime). Rat anti-DE-cadherin (1∶20), mouse anti-Armadillo (1∶20) and mouse anti-Dl 9B (1∶20) were provided by Developmental Studies Hybridoma Bank at the University of Iowa. Secondary antibodies: Alexa 488 and Alexa 568 conjugated secondary antibodies (1∶5000; Molecular Probes); Cy5 conjugated secondary antibodies (1∶1000; Jackson ImmunoResearch Laboratories). All images were captured using an Axioplan2 epi-fluorescence microscope equipped with an Axiocam digital camera (Carl Zeiss, Inc.).