Figure 7.
Nanoscopic cytoskeleton in the MO requires short microtubules. (A) Representative confocal images (lateral view) showing GFP-tubulin in the outer segment of leg receptors in wild-type and six mutants. 1: +/uas-gfp-αtub84B; dcx-emap-gal4, uas-cd4-tdTom. 2: dcx-emap-gal4/uas-gfp-αtub84B; c01236/BE6. 3: +/uas-gfp-αtub84B; dcx-emap-gal4, uas-cd4-tdTom/kat-60L1i-32506. 4: uas-gfp-αtub84B/kat-60L1i-36866; +/dcx-emap-gal4, uas-cd4-tdTom. 5: uas-gfp-αtub84B/patronini-108927; +/dcx-emap-gal4, uas-cd4-tdTom. 6: +/uas-gfp-αtub84B; dcx-emap-gal4, uas-cd4-tdTom/patronini-1753. 7: dcx-emap-gal4/uas-gfp-αtub84B; patronini-1753/kat-60L1i-32506. Scale bar, 5 µm. (B) Representative intensity line profiles of GFP-tubulin signal in the outer segment of leg receptors in wild-type and mutant flies. The numbers correspond to those in A. (C) Statistical quantification of GFP-tubulin signal in the TB and MO of leg receptors. The number of flies is indicated in parentheses. Data are presented as mean ± SD with scattered points. For comparison between different genotypes, two-sided unpaired Student’s t test was used. For comparison between TB and MO from the same cell, two-sided paired Student’s t test was used. **, P < 0.01; ***, P < 0.001. n.s., no significance. (D) Representative ET slice images of the outer segment of leg receptors in kat-60L1i-32506 (two lateral views of the same receptor, but at different layers). Red arrowhead, the long microtubules extending from TB into MO. Red asterisk, the large gap between the microtubule and MO membrane. Note that there were more TB microtubules than in c01236/BE6 (right panel). Scale bar, 0.5 µm. (E) Statistical quantification of the ratio between the number of microtubules that spanned across the TB and MO to the number of MO microtubules in the leg receptors in wild type and kat-60L1i-32506. (F) Statistical quantification of microtubule density in the TB (upper) and the width of the microtubule array in the MO (lower) in the leg receptors in wild type, c01236/BE6, and kat-60L1i-32506. In E and F, the number of cells is indicated. Data are presented as mean ± SD with scattered points. Two-sided unpaired Student’s t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. n.s., no significance. (G) Representative confocal images showing the thick microtubule bundles when Patronin was overexpressed (n = 3 cells, dcx-emap-gal4, uas-mcherry-αtub84B/uas-patronin-gfp). Scale bar, 5 µm.

Nanoscopic cytoskeleton in the MO requires short microtubules. (A) Representative confocal images (lateral view) showing GFP-tubulin in the outer segment of leg receptors in wild-type and six mutants. 1: +/uas-gfp-αtub84B; dcx-emap-gal4, uas-cd4-tdTom. 2: dcx-emap-gal4/uas-gfp-αtub84B; c01236/BE6. 3: +/uas-gfp-αtub84B; dcx-emap-gal4, uas-cd4-tdTom/kat-60L1i-32506. 4: uas-gfp-αtub84B/kat-60L1i-36866; +/dcx-emap-gal4, uas-cd4-tdTom. 5: uas-gfp-αtub84B/patronini-108927; +/dcx-emap-gal4, uas-cd4-tdTom. 6: +/uas-gfp-αtub84B; dcx-emap-gal4, uas-cd4-tdTom/patronini-1753. 7: dcx-emap-gal4/uas-gfp-αtub84B; patronini-1753/kat-60L1i-32506. Scale bar, 5 µm. (B) Representative intensity line profiles of GFP-tubulin signal in the outer segment of leg receptors in wild-type and mutant flies. The numbers correspond to those in A. (C) Statistical quantification of GFP-tubulin signal in the TB and MO of leg receptors. The number of flies is indicated in parentheses. Data are presented as mean ± SD with scattered points. For comparison between different genotypes, two-sided unpaired Student’s t test was used. For comparison between TB and MO from the same cell, two-sided paired Student’s t test was used. **, P < 0.01; ***, P < 0.001. n.s., no significance. (D) Representative ET slice images of the outer segment of leg receptors in kat-60L1i-32506 (two lateral views of the same receptor, but at different layers). Red arrowhead, the long microtubules extending from TB into MO. Red asterisk, the large gap between the microtubule and MO membrane. Note that there were more TB microtubules than in c01236/BE6 (right panel). Scale bar, 0.5 µm. (E) Statistical quantification of the ratio between the number of microtubules that spanned across the TB and MO to the number of MO microtubules in the leg receptors in wild type and kat-60L1i-32506. (F) Statistical quantification of microtubule density in the TB (upper) and the width of the microtubule array in the MO (lower) in the leg receptors in wild type, c01236/BE6, and kat-60L1i-32506. In E and F, the number of cells is indicated. Data are presented as mean ± SD with scattered points. Two-sided unpaired Student’s t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. n.s., no significance. (G) Representative confocal images showing the thick microtubule bundles when Patronin was overexpressed (n = 3 cells, dcx-emap-gal4, uas-mcherry-αtub84B/uas-patronin-gfp). Scale bar, 5 µm.

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