Figure - PMC

Skip to main content

View full-text article in PMC

. Author manuscript; available in PMC: 2022 May 1.

Published in final edited form as: Hepatology. 2022 Feb 8;75(5):1110–1122. doi: 10.1002/hep.32298

FIG. 6. — Depletion of ARL13B and ARL3 impacts ciliary localization of a G-protein coupled bile acid receptor TGR5. (A) TGR5 (green) is localized to cholangiocyte cilia (red) in healthy humans but not in patients with ADPKD-associated PLD and (B) in cultured NHC but not in PLDC where it is overexpressed on the apical membrane. Nuclei (blue, DAPI). (C) TGR5 was not detected in isolated PLDC cilia. (D) By PLA, ARL13B and TGR5 (red) and ARL3 and TGR5 (red) interact in cholangiocyte cilia (green) of healthy humans but not in cilia of patients with PLD. (E) Transfection of NHC with doxycycline-inducible ARL13B (NHC^ΔARL13B) reduced ARL13B expression by 2.2-fold. NHC transfected with ARL13B-GFP were immunoprecipitated (IP) for ARL13B using GFP-Trap agarose beads and then immunoblotted (IB) for ARL3 and TGR5 demonstrating their interaction (2.4-fold and 4.1-fold increase, respectively). (F) NHC^ΔARL13B have shortened cilia (green) and TGR5 (red) is absent in cilia. n=3 for each experiment.

FIG. 6. — Depletion of ARL13B and ARL3 impacts ciliary localization of a G-protein coupled bile acid receptor TGR5. (A) TGR5 (green) is localized to cholangiocyte cilia (red) in healthy humans but not in patients with ADPKD-associated PLD and (B) in cultured NHC but not in PLDC where it is overexpressed on the apical membrane. Nuclei (blue, DAPI). (C) TGR5 was not detected in isolated PLDC cilia. (D) By PLA, ARL13B and TGR5 (red) and ARL3 and TGR5 (red) interact in cholangiocyte cilia (green) of healthy humans but not in cilia of patients with PLD. (E) Transfection of NHC with doxycycline-inducible ARL13B (NHC^ΔARL13B) reduced ARL13B expression by 2.2-fold. NHC transfected with ARL13B-GFP were immunoprecipitated (IP) for ARL13B using GFP-Trap agarose beads and then immunoblotted (IB) for ARL3 and TGR5 demonstrating their interaction (2.4-fold and 4.1-fold increase, respectively). (F) NHC^ΔARL13B have shortened cilia (green) and TGR5 (red) is absent in cilia. n=3 for each experiment.