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Figure 2:Wurst-mediated airway clearance is tracheal specific and independent of posterior spiracles Bright-field in vivo microscopy (A,B,F,H) of late embryos at stage 17. Red dashes indicate air-filled, blue dashes liquid-filled airways. Confocal (C,D) and in vivo fluorescence microscopy (E,G) images of stage 16 embryos. (A) Wild-type embryos undergo air-filling (B) wurstl(1)G0162 mutant embryos lack air-filling (C) RNA in situ hybridization reveals wurst transcript expression in the foregut (fg), hindgut (hg), epidermis (ep), tracheal system (ts) and posterior spiracles (psp). (D) Wurst and RFP were specifically expressed under the control of the UASGAL4 system. Immunofluorescence studies using anti-Wurst and anti-RFP antibodies reveal ectopic Wurst and RFP co-expression (orange, arrowheads) in the characteristic stripe pattern of the paired GAL4 driver line. Note the additional endogenous Wurst (green) expression in tracheal cells and posterior spiracles (arrows). (E,F) The failure of air-filling in wurstl(1)G0162 mutants can be rescued by ectopically expressing wurst in tracheal cells, using the tracheal specific btlG4 driver line. Arrowheads in E point to RFP expression in the tracheal system (ts). (G,H) The failure of air-filling in wurstl(1)G0162 mutants cannot be rescued by ectopically expressing wurst in posterior spiracle (psp) cells, using the dppG4 driver line. Arrowheads in G point to the RFP expression in psp.
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