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. 2020 Dec 28;4(3):e202000746. doi: 10.26508/lsa.202000746

Figure 6. HTR6 CTS1 and CTS2 are both sufficient for ciliary targeting.

Figure 6.

(A) Schematic of HTR7 with or without the CTS2-containing C-terminal tail of HTR6 (HTR6-CT) fused to its C terminus. (A, B) IMCD3 cells expressing C-terminally EGFP-tagged constructs from (A) were analyzed by immunofluorescence with antibodies against EGFP (green), acetylated tubulin (AcTub, red) and gamma-tubulin (γTub, blue). Scale bar, 5 μm. (B, C) Percentage of G protein-coupled receptor-positive cilia relative to total transfected-cell cilia was quantitated from (B). (D) Schematic of CD8α(1-206) chimeras, containing extracellular and transmembrane regions of CD8α fused to HTR7-CT, HTR6-CT (containing CTS2) or HTR6-IC3 (containing CTS1). (B, D, E) IMCD3 cells expressing C-terminally EYFP-tagged constructs from (D) were analyzed as in (B). Scale bar, 5 μm. (E, F) Percentage of G protein–coupled receptor-positive cilia relative to total transfected-cell cilia was quantitated from (E). Data in (C, F) are mean ± SEM of n = 4–5 independent experiments per construct. In each experiment, at least 50 cilia were counted per condition. (C, F) Data were analyzed by unpaired two-tailed t test (C) or by one-way ANOVA followed by Tukey’s multiple comparison tests (F). Significance in both cases is shown as P < 0.0001 (****).