Fig. 5: Dysfunction of peroxisomal metabolism alters cholesterol distribution in the cells and modifies Yap signaling. | Cell Death & Disease

Fig. 5: Dysfunction of peroxisomal metabolism alters cholesterol distribution in the cells and modifies Yap signaling.

From: Peroxisomal cholesterol metabolism regulates yap-signaling, which maintains intestinal epithelial barrier function and is altered in Crohn’s disease

Fig. 5

A Imaging of Filipin III staining to visualize cholesterol in WT and Pex2−/− MIO-derived monolayers. The bar graph represents the size of Filipin III puncta (highlighted by the red arrows), n = 40 monolayers derived from four mice per genotype. Scalebar = 10 µm. B Immunofluorescence with Filipin III in WT and Pex2−/− MIO-derived monolayer untreated and treated with 10 mM MβCD for 2 h. The bar graph represents the size of cholesterol puncta, n = 40 monolayers images derived from four established monolayers from four mice per genotype. Scalebar = 10 µm. C IF image of p-Yap in MIO-derived monolayer, untreated and treated with 10 mM MβCD for 2 h; The bar graph reports the percentage of p-Yap fluorescent signal that overlaps with the DAPI fluorescent signal. n = 30 monolayers derived from three established monolayers from three mice per genotype. D Imaging of FIlipin III staining in and Mex > w1118 and Mex > Pex5-i enterocytes; n = 20 intestines per genotype. Scalebar = 10 µm. E Representative pictures of Mex > w1118 and Mex > Pex5-i flies fed a regular cornmeal diet or a diet supplemented with 10 mM MβCD for 48 h before being tested in a Smurf assay. Scalebar = 500 µm. The dotted lines in the right panels are enterocyte boundary traces of the above panels to illustrate enterocyte size. The bar graph indicates the percentage of dark blue area relative to the total abdomen area; n = 20 guts per genotype and condition. F Representative pictures of flies of the reported genotypes tested in the Smurf assay. The dotted lines in the right panels are enterocyte boundary traces of the above panels to illustrate enterocyte size. The bar graph indicates the percentage of dark blue area in the total abdomen area; n = 20 guts per genotype. Scalebar = 500 µm. In all histograms, the error bars represent standard deviations. Significance was determined using Student’s t-test in A and D and a two-way ANOVA test for all the other graphs. ****p < 0.0001; ***p < 0.001; **p < 0.01; ns not significant.

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