Figure 3.
DCX-EMAP is a structural component of the microtubule-based cytoskeleton in the MOs. (A) Endogenous localization of GFP-DCX-EMAP (DCX-EMAPKI) in haltere campaniform receptors (top view). Scale bar, 10 µm. Inset, localization of GFP-DCX-EMAP in the outer segment of a haltere campaniform receptor (lateral view). Inset scale bar, 2 µm. Genotype: DCX-EMAP-gal4, uas-cd4-tdtom, and DCX-EMAPKI. (B) A representative airyscan superresolution image out of all data from four flies and in total 65 cells (upper) showing the localization of GFP-DCX-EMAP (DCX-EMAPKI) at the central region of the MOs (haltere receptor, top view). The corresponding intensity line profiles of the membrane (red) and GFP-DCX-EMAP (green) are shown in the lower panel. Scale bar, 1 µm. (C) Endogenous localization of GFP-DCX-EMAP (DCX-EMAPKI) in leg campaniform (upper panel) and labellum bristle (lower panel, lateral view) receptors. Genotype: DCX-EMAP-gal4, uas-cd4-tdtom, DCX-EMAPKI. Scale bar, 10 µm. (D) Localization of GFP-DCX-EMAP in haltere (top view) and leg (lateral view) receptors in DCX-EMAPOE strain. Genotype: DCX-EMAP-gal4, uas-cd4-tdtom/uas-gfp-DCX-EMAP. Scale bar, 10 µm. (E) Three sets of representative confocal images (lateral view of the leg receptors) showing the recovery of GFP-DCX-EMAP (DCX-EMAPKI) signal, mCherry-tubulin (DCX-EMAP-gal4/uas-mcherry-αTub84B), and EB1-GFP (DCX-EMAP-gal4; uas-Eb1-gfp). −1 s, right before bleaching. 0 s, just after bleaching. 30 or 15 min, time after bleaching. The white arrowhead in each panel indicates the position of the MO. Scale bar, 5 µm. (F) Fluorescence recovery curves after photobleaching. Green (open circle), GFP-DCX-EMAP (DCX-EMAPKI) recovery in the MOs (n = 6 cells). Red (open circle), mCherry-tubulin recovery in the outer segments (n = 6 cells). Black (solid circle), EB1-GFP recovery in the outer segments (n = 12 cells). Data are presented as mean ± SEM. (G) Representative images (lateral view) showing the localization of GFP-DCX-EMAP in the leg receptor of wild type (DCX-EMAP-gal4, uas-cd4-tdtom/uas-gfp-DCX-EMAP) or the kat-60L1 mutant (DCX-EMAP-gal4/uas-gfp-DCX-EMAP; c12306/BE6). Three types of phenotypes were identified, and the number of cells falling into each type was indicated. Scale bar, 10 µm. (H and I) Representative images showing the localization of GFP-DCX-EMAP in the haltere (top view; H) and leg receptors (lateral view; I) of wild type (DCX-EMAPKI/DCX-EMAP-gal4,uas-cd4-tdtom) or the nompC null mutant (nompC3; DCX-EMAPKI/DCX-EMAP-gal4,uas-cd4-tdtom). Scale bar, 5 µm. (J) Statistical quantification of DCX-EMAP signal in the MOs of haltere (n = 3 halteres for wild type and nompC3, respectively) and leg receptors (wild type: n = 12 cells; nompC3: n = 10 cells). Data are presented as mean ± SD with scattered data points. Two-sided unpaired Student’s t test. n.s., no significance.

DCX-EMAP is a structural component of the microtubule-based cytoskeleton in the MOs. (A) Endogenous localization of GFP-DCX-EMAP (DCX-EMAPKI) in haltere campaniform receptors (top view). Scale bar, 10 µm. Inset, localization of GFP-DCX-EMAP in the outer segment of a haltere campaniform receptor (lateral view). Inset scale bar, 2 µm. Genotype: DCX-EMAP-gal4, uas-cd4-tdtom, and DCX-EMAPKI. (B) A representative airyscan superresolution image out of all data from four flies and in total 65 cells (upper) showing the localization of GFP-DCX-EMAP (DCX-EMAPKI) at the central region of the MOs (haltere receptor, top view). The corresponding intensity line profiles of the membrane (red) and GFP-DCX-EMAP (green) are shown in the lower panel. Scale bar, 1 µm. (C) Endogenous localization of GFP-DCX-EMAP (DCX-EMAPKI) in leg campaniform (upper panel) and labellum bristle (lower panel, lateral view) receptors. Genotype: DCX-EMAP-gal4, uas-cd4-tdtom, DCX-EMAPKI. Scale bar, 10 µm. (D) Localization of GFP-DCX-EMAP in haltere (top view) and leg (lateral view) receptors in DCX-EMAPOE strain. Genotype: DCX-EMAP-gal4, uas-cd4-tdtom/uas-gfp-DCX-EMAP. Scale bar, 10 µm. (E) Three sets of representative confocal images (lateral view of the leg receptors) showing the recovery of GFP-DCX-EMAP (DCX-EMAPKI) signal, mCherry-tubulin (DCX-EMAP-gal4/uas-mcherry-αTub84B), and EB1-GFP (DCX-EMAP-gal4; uas-Eb1-gfp). −1 s, right before bleaching. 0 s, just after bleaching. 30 or 15 min, time after bleaching. The white arrowhead in each panel indicates the position of the MO. Scale bar, 5 µm. (F) Fluorescence recovery curves after photobleaching. Green (open circle), GFP-DCX-EMAP (DCX-EMAPKI) recovery in the MOs (n = 6 cells). Red (open circle), mCherry-tubulin recovery in the outer segments (n = 6 cells). Black (solid circle), EB1-GFP recovery in the outer segments (n = 12 cells). Data are presented as mean ± SEM. (G) Representative images (lateral view) showing the localization of GFP-DCX-EMAP in the leg receptor of wild type (DCX-EMAP-gal4, uas-cd4-tdtom/uas-gfp-DCX-EMAP) or the kat-60L1 mutant (DCX-EMAP-gal4/uas-gfp-DCX-EMAP; c12306/BE6). Three types of phenotypes were identified, and the number of cells falling into each type was indicated. Scale bar, 10 µm. (H and I) Representative images showing the localization of GFP-DCX-EMAP in the haltere (top view; H) and leg receptors (lateral view; I) of wild type (DCX-EMAPKI/DCX-EMAP-gal4,uas-cd4-tdtom) or the nompC null mutant (nompC3; DCX-EMAPKI/DCX-EMAP-gal4,uas-cd4-tdtom). Scale bar, 5 µm. (J) Statistical quantification of DCX-EMAP signal in the MOs of haltere (n = 3 halteres for wild type and nompC3, respectively) and leg receptors (wild type: n = 12 cells; nompC3: n = 10 cells). Data are presented as mean ± SD with scattered data points. Two-sided unpaired Student’s t test. n.s., no significance.

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