Fig. 4

5-HT6 mutations regulated the locations of ARL13B and AnkG proteins. (a) D72A, D106A, and F69L + T70I + D72A mutations influenced the ciliary location of ARL13B. Overexpression of 5-HT6 delocalized ARL13B in primary cilia. D72A, D106A, and F69L + T70I + D72A mutations of 5-HT6 led to relocation of ARL13B (red) into the cilia. 5-HT6 (green) indicated primary cilia of hippocampal neurons. (b) D72A, D106A, and F69L + T70I + D72A mutations restored the ciliary location of ARL13B. Overexpression of 5-HT6 led to delocalization of ARL13B in primary cilia. EGFP, ***p < 0.001; D72A, *p < 0.05; D106A, *p < 0.05; F69L + T70I + D72A, *p < 0.05 (Kruskal–Wallis test, p < 0.001). (c) AnkG was originally located at the AIS, but it was located at the primary cilia following overexpression of 5-HT6. 5-HT6 mutations did not restore the location of AnkG. EGFP, ***p < 0.001 (Kruskal–Wallis test, p < 0.001). All groups were compared with the 5-HT6 group, which was transfected with 5-HT6-GFP. Scale bars, 10 μm. All data presented as mean ± SEM (≥3 independent experiments). 5-HT6, serotonin 6 receptor, DAPI 4',6-diamidino-2-phenylindole, EGFP enhanced green fluorescent protein