A, B) Representative images of EBP-2::GFP (A) or RFP::PTRN-1 (B) localization in the AWA cilia of animals of the indicated genotypes. Insets in B show localization patterns in the axonemes; images have been overexposed to allow visualization of the faint puncta. Both fusion proteins were expressed under the gpa-4Δ6 promoter. Numbers at top right indicate the percentage of neurons exhibiting the phenotype; n≥30 each in 3 independent experiments. The cilia base and cilia are indicated by yellow and white arrowheads, respectively; the dendrite is marked by an arrow. AWA neurons were visualized via expression of gpa-4Δ6p::myr-gfp in B. Anterior is at top or at top left in all images. Scale bars: 10 μm.
C) Representative images of TBB-4::GFP localization in wild-type and dyf-18(ok200) mutants. TBB-4::GFP was expressed under the gpa-4Δ6 promoter. AWA cilia were visualized via expression of gpa-4Δ6p::myr-mCherry. Numbers at top right indicate the percentage of animals exhibiting the phenotype; n≥30 neurons each in 3 independent experiments. The cilia base and cilia are indicated by yellow and white arrowheads, respectively; the dendrite is marked by an arrow. Anterior is at left or at top left in all images. Scale bar: 10 μm.
D) (Top) Representative images of AWA cilia pre-bleach (Pre), at bleach (0h) and 12h post-bleaching (12h). The bleached area is indicated by a dotted box. Note that while the same animal was examined at all three timepoints, the animals were removed after bleaching to growth plates, and re-examined after 12h (see STAR Methods). Cilia are indicated by white arrowheads; the dendrite is marked by an arrow. Anterior is at top. Scale bar: 5 μm. (Bottom) Mean fluorescence intensity ratios (photobleached to unbleached regions within an AWA cilium; see STAR Methods) across a line segment on AWA cilia branches shown at 0 hour and 12 hour post-bleaching for individual wild-type and dyf-18(ok200) animals. Statistical significances were calculated using a paired t-test. Experiments were performed on 3 independent days.
Also see Figure S3.