Figure S1.
KI strains and live-cell imaging of leg campaniform mechanoreceptors. (A) Cartoon schematic for the NompC-GFP KI strain (nompC-gfp-KI). The insertion site of GFP was indicated. (B) Localization of NompC-GFP (nompC-gfp-KI) in fly bristle mechanoreceptors (left panel) and haltere campaniform mechanoreceptors (middle and right panels). These observations were consistent with the immunostaining data obtained using a NompC monoclonal antibody (Liang et al., 2011). Scale bar, 20 µm. (C) A decapitated fly was kept in a wet chamber, and the locations of leg campaniform mechanoreceptors are indicated (red arrowheads). During imaging, the fly was immobilized on a cover glass using double-sided tape. The three legs on the same body side were covered by the second coverslip to facilitate imaging using an inverted spinning-disk confocal microscope. s3, the third segment. s5, the fifth segment. (D and E) Campaniform mechanoreceptors at the third (s3) and fifth (s5) segments of a fly leg (genotype, dcx-emap-gal4/uas-cd4-tdTom). The outer segment in these cells is indicated by a white arrow. Scale bar, 5 µm. (F) Cartoon schematic for the Patronin-RFP KI strain (patronin-rfp-KI). The insertion site of RFP is indicated. Based on the genome annotation in Flybase, all Patronin isoforms were tagged. (G) Using an endogenously tagged Shot-GFP (Sun et al., 2019b) as a marker, we showed that Shot-GFP and Patronin-RFP together formed puncta signals in epidermal cells (left and middle panels) and campaniform mechanoreceptors (right panel) in fly legs (genotype, Shot-GFP/patronin-rfp-KI). In these puncta, Shot and Patronin showed either contiguous or partially overlapping localizations (white arrowheads, two regions enlarged in the insets, right panel), consistent with the Shot-Patronin foci observed in the fly embryo (Nashchekin et al., 2016). Yellow arrowheads (right) indicate two dot-shaped Shot-Patronin signals that were in the thecogen cell. Scale bars, 1 µm. (H) Cross-sectional view (ET slice image) of the cilium and its surrounding structures in a haltere receptor. Scale bar, 250 nm. (I) Lateral view (ET slice image) of the structures near the cilium in a leg receptor. Scale bar, 250 nm. In H and I, microtubules in the thecogen cells are indicated by red arrowheads.

KI strains and live-cell imaging of leg campaniform mechanoreceptors. (A) Cartoon schematic for the NompC-GFP KI strain (nompC-gfp-KI). The insertion site of GFP was indicated. (B) Localization of NompC-GFP (nompC-gfp-KI) in fly bristle mechanoreceptors (left panel) and haltere campaniform mechanoreceptors (middle and right panels). These observations were consistent with the immunostaining data obtained using a NompC monoclonal antibody (Liang et al., 2011). Scale bar, 20 µm. (C) A decapitated fly was kept in a wet chamber, and the locations of leg campaniform mechanoreceptors are indicated (red arrowheads). During imaging, the fly was immobilized on a cover glass using double-sided tape. The three legs on the same body side were covered by the second coverslip to facilitate imaging using an inverted spinning-disk confocal microscope. s3, the third segment. s5, the fifth segment. (D and E) Campaniform mechanoreceptors at the third (s3) and fifth (s5) segments of a fly leg (genotype, dcx-emap-gal4/uas-cd4-tdTom). The outer segment in these cells is indicated by a white arrow. Scale bar, 5 µm. (F) Cartoon schematic for the Patronin-RFP KI strain (patronin-rfp-KI). The insertion site of RFP is indicated. Based on the genome annotation in Flybase, all Patronin isoforms were tagged. (G) Using an endogenously tagged Shot-GFP (Sun et al., 2019b) as a marker, we showed that Shot-GFP and Patronin-RFP together formed puncta signals in epidermal cells (left and middle panels) and campaniform mechanoreceptors (right panel) in fly legs (genotype, Shot-GFP/patronin-rfp-KI). In these puncta, Shot and Patronin showed either contiguous or partially overlapping localizations (white arrowheads, two regions enlarged in the insets, right panel), consistent with the Shot-Patronin foci observed in the fly embryo (Nashchekin et al., 2016). Yellow arrowheads (right) indicate two dot-shaped Shot-Patronin signals that were in the thecogen cell. Scale bars, 1 µm. (H) Cross-sectional view (ET slice image) of the cilium and its surrounding structures in a haltere receptor. Scale bar, 250 nm. (I) Lateral view (ET slice image) of the structures near the cilium in a leg receptor. Scale bar, 250 nm. In H and I, microtubules in the thecogen cells are indicated by red arrowheads.

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