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. 2016 Jun 1;6:27119. doi: 10.1038/srep27119

Figure 3. Isolation of villi and crypt epithelial cells of the jejunum from BL-6 WT and PepT1 KO mice.

Figure 3

(a) Total RNAs were extracted from the different fractions collected from BL-6 WT mice using the low-temperature method, and the expression levels of Muc2 (as a villus marker) and Lgr5 (as a crypt marker) were assessed by qRT-PCR. (b) Total RNAs were extracted from different fractions collected from PepT1 KO mice, and the expression levels of Muc2 (as a villus marker) and Lgr5 (as a crypt marker) were assessed by qRT-PCR. (c) Pictures of selected villi and crypts fractions were taken with a Nikon Eclipse TS100 microscope at 10× magnifications. Scale bar, 50 μm. (d) Total RNAs were extracted from selected villus and crypt fractions from BL-6 WT and PepT1 KO mice. The expression levels of Muc2, Lgr5 and mPepT1 in the villi (blue bars) and crypts (magenta) of BL-6 WT and PepT1 KO mice were further assessed by qRT-PCR (n = 5; and **P < 0.005, ***P < 0.001). (e) The expression of Muc2, Lgr5 and mPepT1 in the tissue were confirmed by immunofluorescence. Muc2, Lgr5, mPepT1 were immunostained using anti-Muc2, anti-Lgr5 and anti-PepT1 (FITC, green), respectively, F-actin was stained using phalloidin (TRITC, red), and cell nuclei were stained using DAPI (blue). Separate pictures were taken at 20× for each filter, and the images were merged. Scale bar, 50 μm.