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. 2021 Mar 17;12:1703. doi: 10.1038/s41467-021-21882-1

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© The Author(s) 2021

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. 1 — a Representative confocal images of YAP/TAZ endogenous immunostaining in MCF10A cells exposed to control, ATG16L1 or ATG7/10 siRNAs. DAPI = nucleus. Scale bars are 10 µm. The experiment was repeated twice with similar results. b YAP/TAZ localisation (nuclear—Nc or cytoplasmic—Cyt) in MCF10A cells exposed to control, ATG16L1 or ATG7/10 siRNAs (n = 126 (Ctrl), 170 (ATG16L1 KD), 166(ATG7/10 KD) cells; ****P < 0.0001; chi-squared test). The experiment was repeated twice with similar results. c Luciferase assay for YAP/TAZ activity in MCF10A cells exposed to control, ATG16L1 or ATG7/10 siRNAs. Bars represent the mean ± s.d. (n = 3 independent experiments; **P < 0.01, *P < 0.05; two-tailed one sample t-test). d Representative confocal images of endogenous YAP/TAZ and F-actin (Phalloidin) in primary mammary epithelial cells (pMECs) isolated from wild-type and Atg16L1 hypomorph (Atg16hyp) mice. Scale bars are 10 µm. The experiment was repeated twice with similar results. e YAP/TAZ localisation in wild-type (n = 3 mice) and Atg16hyp (n = 4 mice) pMECs. Bars represent the mean ± s.d. (****P < 0.0001; two-way ANOVA). f Luciferase assay for YAP/TAZ activity in wild-type and Atg16hyp pMECs. Bars represent the mean ± s.d. (n = 3 mice; **P < 0.01; two-tailed one sample t-test). g Percentage of cells with F-actin stress fibres in wild-type and Atg16hyp pMECs (n = 98 (wild-type) and 292 (Atg16hyp) cells; ****P < 0.0001; chi-squared test). The experiment was repeated twice with similar results. h Representative images of BrdU immunostaining in MCF10A cells exposed to control, ATG16L1 or ATG7/10 siRNAs. Scale bars are 10 µm. The experiment was repeated twice with similar results. i The graphs show the percentages of BrdU-positive MCF10A cells, after exposure to control, ATG16L1 or ATG7/10 siRNAs: mean ± s.d. (n = 3 independent experiments; ****P < 0.0001, **P < 0.01; two-tailed t-test). j Size of autophagy inhibited cells: MCF10A cells exposed to control, ATG16L1 (n = 3 independent experiments) or ATG7/10 (n = 5 independent experiments) siRNAs. Confocal images of each cell type were analysed to measure cell area using ZEN software. Bars represent the mean ± s.d. (****P < 0.0001, **P < 0.01; two-tailed t-test). Exact P values for asterisks: c (from left to right) 0.0132, 0.0010; f 0.0033; i 0.0058; and j 0.0061.