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. 2010 Dec 1;21(23):4141–4150. doi: 10.1091/mbc.E10-05-0424

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© 2010 by The American Society for Cell Biology

This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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Figure 9. — Knockdown of PI4KIIα or PI4KIIIβ does not prevent oxysterol-dependent Golgi localization of OSBP. CHO cells transfected with siNT (A and C), siPI4KIIα (B), or siPI4KIIIβ (D) were treated with ethanol solvent or 25-hydroxycholesterol (25OH; 2.5 μg/ml) for 1 h. Cells were then fixed and incubated with an OSBP polyclonal antibody and goat anti-rabbit Alexa Fluor488 (C and D) or Alexa Fluor594 (A and B) secondary antibodies, followed by PI4KIIα or PI4KIIIβ monoclonal antibodies and a goat anti-mouse Alexa Fluor594 C and D) or Alexa Fluor488 (A and B) secondary antibodies. Images are single confocal sections (0.4–0.2 μM) taken on an LSM510/AxioVert 100M inverted microscope equipped with a 100× oil immersion objective (NA 1.4). HeLa cells were transiently transfected with siPI4KIIα (E) or siPI4KIIIβ (F), treated with 25-hydroxycholesterol for 1 h, and immunostained for the corresponding PI4K and OSBP. Images were captured on an Axiovert 200M fluorescence microscope equipped with a 63× objective (NA 1.4).