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. 2024 Apr 25;15:3365. doi: 10.1038/s41467-024-47244-1

Fig. 1. Cilium-TurboID selectively biotinylates ciliary proteins.

Fig. 1

a Schematics of the truncated Arl13b constructs used. Y and N indicate whether the corresponding construct localizes to the cilium or not, respectively. Y/N: this construct localizes to both the cilium and the cytosol. “+”: the degree of cilium elongation in NIH3T3 cells transiently transfected with the corresponding constructs. “-”: cilium length is not impacted. b Immunofluorescence of stable NIH3T3 cell lines expressing cilium-TurboID (N + RVEP-PR-EGFP-TurboID) or non-cilium-TurboID (PR-EGFP-TurboID) before and after biotin labeling. Cells were serum-starved for 24 h, incubated with 50 μΜ biotin for 10 min at 37 °C, and fixed for immunostaining. The cilium is labeled by immunostaining for acetylated-tubulin (red, ac-tubulin), and the biotinylated proteins were detected via streptavidin-Alexa Fluor 647 (magenta). Cilium-TurboID and non-cilium TurboID were detected by immunostaining for EGFP (green). Scale bars, 10 µm in all panels. Experiments were performed three times with similar results.