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. 2017 Jan 23;36(24):3450–3463. doi: 10.1038/onc.2016.495

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Copyright © 2017 The Author(s)

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/4.0/

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AURKA and HDAC6 inhibition rescues ciliogenesis in VHL-deficient cells. (a) Representative images from hTERT RPE-1 cells transiently transfected with siControl (siC) or siVHL, treated with vehicle (DMSO), alisertib (MLN8237) or rocilinostat (ACY1215) at the time of serum withdrawal for 48 h. Ciliation monitored by immunofluorescent staining using acetylated α-tubulin (cilia marker) and pericentrin (basal body marker). Nuclei counterstained using DAPI. Highlighted boxes show magnified cilia. Scale bar, 3 μM. (b, d) Percentage of cells that fail to ciliate (unciliated) normalized to 1, from siC or siVHL transfected cells treated with (b) alisertib or (d) rocilinostat. * and ^# denote statistical significance. (c, e) Cilia length (μM) from siC or siVHL transfected cells treated with (c) alisertib or (e) rocilinostat. At least 100 cells were counted from each replicate. * and ^# denote statistical significance. All error bars indicate s.e.m. (f) Model depicting a role for VHL’s ubiquitin ligase function in conditions of normoxia targeting HIFα and hypoxia targeting AURKA.

AURKA and HDAC6 inhibition rescues ciliogenesis in VHL-deficient cells. (a) Representative images from hTERT RPE-1 cells transiently transfected with siControl (siC) or siVHL, treated with vehicle (DMSO), alisertib (MLN8237) or rocilinostat (ACY1215) at the time of serum withdrawal for 48 h. Ciliation monitored by immunofluorescent staining using acetylated α-tubulin (cilia marker) and pericentrin (basal body marker). Nuclei counterstained using DAPI. Highlighted boxes show magnified cilia. Scale bar, 3 μM. (b, d) Percentage of cells that fail to ciliate (unciliated) normalized to 1, from siC or siVHL transfected cells treated with (b) alisertib or (d) rocilinostat. * and ^# denote statistical significance. (c, e) Cilia length (μM) from siC or siVHL transfected cells treated with (c) alisertib or (e) rocilinostat. At least 100 cells were counted from each replicate. * and ^# denote statistical significance. All error bars indicate s.e.m. (f) Model depicting a role for VHL’s ubiquitin ligase function in conditions of normoxia targeting HIFα and hypoxia targeting AURKA.