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. 2023 Jan 20;12:e80135. doi: 10.7554/eLife.80135

Figure 4. 3D cyst cultures demonstrate disrupted epithelial polarity.

Figure 4.

(A) 3D cyst assay were performed with WT and KO cultures. Immunofluorescence micrographs of 3D cysts generated from WT and KO cell lines. All knockdown cell lines form multiple lumina or no lumina. DPP4 localizes to actin-rich structures in al KO cell lines and additionally, to intracellular, actin-negative compartments in TM9SF4-, MTMR2- and ANO8-KO clones (white arrowheads). Scale = 10 µm. (B) Single central lumen formation was quantified. The percentage of cysts with a single central lumen is substantially decreased in the respective KO cells lines (dot box plot, Mann–Whitney U test. ***p< 0.005, n ≥ 100 cells per condition). (C) Delocalized DPP4 in cysts was quantified (dot box plot, Mann–Whitney U test. ***p<0.005, n.s., not significant).