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. 2019 Feb 21;38(24):4739–4754. doi: 10.1038/s41388-019-0754-1

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — All CCT subunits are required for the CCT complex function. a CCT4 staining of ptc>CCT4 RNAi wing disc. ptc>CCT4 RNAi caused reduction in CCT4 staining in ptc-expressing area (GFP positive). Scale bar, 50 μm. b High magnification views of the box area in a. Scale bar, 25 μm. c CCT4 staining in wing discs with CCT subunit RNAi. nub>+ shows control wing pouch with even CCT staining. The nub-expressing area is marked by red dashed line. CCT4 staining in nub-expressing area was reduced when CCT1–8 was knocked down. mCherry expression by nub-Gal4 is shown in c′. Green staining indicates phalloidin. Scale bar, 100 μm. d Effects of CCT subunit RNAi on the CCT complex. CCT1 and CCT4 levels were decreased by reducing any of eight CCT subunits in S2R+ cells. GAPDH levels were used as a loading control. e Western blot of wild type and CCT4^KG09280/+ adult extracts. The level of CCT4 was decreased in CCT4^KG09280/+ extracts compared to wild-type control. Note that CCT1 was also reduced by the CCT4 mutation. Histone 3 and GAPDH were used as loading controls. f All CCT subunit RNAi in ey-Gal4 resulted in late pupal lethality with similar headless phenotypes