Figure 1.
Figure 1. SNAP-tagged gp135 traffics to the apical membrane and is delivered to the periciliary ring. (A) Schematic view of the SNAP-tagged gp135 construct and BG labeling. DTHL is the amino acid sequence of the C termins of the gp135 protein. (B) MDCK cells stably expressing SNAP-gp135 were fixed, stained with BG549 (red), and processed for immunofluorescence (ZO1, green; acetylated tubulin, blue). Arrowheads indicate positions of the orthogonal views. Arrows indicate ciliary exclusion zones. (C–E) Old surface SNAP-gp135 was labeled to saturation with BG549 (red). Newly delivered SNAP-gp135 was labeled with BG488 (green) after the indicated times at 37°C (ZO1, blue). Arrowheads indicate periciliary rings. (D and E) 3D reconstructions demonstrating SNAP-gp135 periciliary delivery. (D) Time = 8 min (acetylated tubulin, magenta). Bars, 10 µm.

SNAP-tagged gp135 traffics to the apical membrane and is delivered to the periciliary ring. (A) Schematic view of the SNAP-tagged gp135 construct and BG labeling. DTHL is the amino acid sequence of the C termins of the gp135 protein. (B) MDCK cells stably expressing SNAP-gp135 were fixed, stained with BG549 (red), and processed for immunofluorescence (ZO1, green; acetylated tubulin, blue). Arrowheads indicate positions of the orthogonal views. Arrows indicate ciliary exclusion zones. (C–E) Old surface SNAP-gp135 was labeled to saturation with BG549 (red). Newly delivered SNAP-gp135 was labeled with BG488 (green) after the indicated times at 37°C (ZO1, blue). Arrowheads indicate periciliary rings. (D and E) 3D reconstructions demonstrating SNAP-gp135 periciliary delivery. (D) Time = 8 min (acetylated tubulin, magenta). Bars, 10 µm.

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