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. 2021 Mar 31;17(3):e1009476. doi: 10.1371/journal.ppat.1009476

Fig 7. Cytoskeleton manipulation partially rescues goblet cell maturation in LSD1-deficient organoids.

Fig 7

(A) cKO organoids were left untreated (ENR) or treated with Y-27632 (10 μM) for 4 days. Representative bright field images are shown. Scale bar is 100 μm. (B) Quantification of organoid area from minimal projections 4 days after seeding. Dots represents mean of one well, pooled from 3 independent experiments. (C&D) Representative immunofluorescent images of MUC2 (green) and β-catenin (purple). MUC2 fluorescence intensity relative to organoid area was measured. Scale bar is 50 μm. (E) cKO organoids were treated with IL-22 (5ng/ml) or IL22+ Y-27632) for 4 days. Representative bright field images are shown. Scale bar is 100 μm. (F) Mean organoid area of cKO organoids is depicted as well as the percentage of spheroids in each well 4 days after seeding. (G) cKO organoids were left untreated (ENR) or treated with IL-22 (5ng/ml), Y-27632 (10 μM) and combination of Y-27632 and IL-22 for 4 days and representative images for RELMβ (green) and β-catenin (purple)staining are shown. (H) Pooled quantification of RELMβ+ cells per organoid area from independent experiments. (I) WT organoids were pre-treated with GSK-LSD1 for 7 days. Next, these organoids were treated with combination of IL-22 (5ng/ml) + GSK-LSD1 and GSK-LSD1+ IL-22+ Y-27632 for 4 days. Immunofluorescence images of RELMβ (green) and β-catenin (purple) are shown. (J) Quantification of RELMβ+ cells per organoid area is shown. Unpaired two-tailed Student’s t test (B, D, F, H & J) was performed to define significance. * P ≤ 0.05, **, P < 0.01, *** P ≤ 0.001.