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. 2021 Aug 4;12:4697. doi: 10.1038/s41467-021-24993-x

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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. 7 — a Schematic diagram showing mutation sites at intracellular and peptidase domain of CD13. b Confocal images of CD13-V5 (magenta) and Par6 (green) showing the rescue phenotype in wildtype and different CD13 mutants of shScr and shCD13 Caco-2 cysts. c Quantification of the percentage of Caco-2 spheroids with internally localized Par6 in wildtype (shScr, n = 515; shCD13, n = 292), CD13-Y6F (shScr, n = 462; shCD13, n = 433), CD13-S8A (shScr, n = 541; shCD13, n = 333), and CD13-H388A (shScr, n = 452; shCD13, n = 336) of shScr (n = 428) and shCD13 (n = 492) Caco-2 cysts. Dots represent mean values ± SD from each of 10 experiments. p-values were calculated by ANOVA with Tukey’s posthoc test for multiple comparisons. d Images for CD13-V5 (magenta) and Par6 (green) showing the rescue phenotype in wildtype and CD13-Y6F mutant of shScr and shCD13 Caco-2 in 2-cell structures. Numbers in the lower right indicate the fraction of cells analyzed with Par6 localized between the nuclei. e Caco-2 cells expressing CD13-WT or CD13-Y6F were immunostained for CD13-V5 (magenta) and Calreticulin to label the endoplasmic reticulum (ER) (green). f Quantification of the proportion of CD13 puncta that overlap with sites of ER-positivity. Dots represent individual Caco-2 structures from a single experiment. p-values were calculated with an unpaired two-tailed student’s t-test. Images are representative from two (b, d) independent experiments. Bars: b, 50 μm; d, e, 10 μm.