Fig. 3: DR5 is needed for the proliferation and differentiation of ISC in the ileum.

Data were expressed as mean ± SEM. Statistical analyses were performed by the unpaired t-tests (*P < 0.05, **P < 0.01, ***P < 0.001). A qRT-PCR analysis to show the expression of ISC marker genes in ileum of WT and DR5^-/- mice (n = 6). B Representative images for immunohistochemical staining of Olfm4 within two groups (scale bars: 100 μm). C Statistical graph of the Olfm4⁺ ISCs number in WT mice and DR5^-/- mice (n = 6). At least 15 crypts were counted randomly for each slide and the mean value was calculated. D Representative images for immunohistochemical staining of Ki67 (scale bars: 100 μm) and quantification of Ki67⁺ cells in ileal crypt in WT mice and DR5^-/- mice (n = 6). At least 15 crypts were counted randomly for each slide and the mean value was calculated. E Representative images for immunohistochemical staining of BrdU (scale bars: 100 μm) and quantification of BrdU⁺ cells in ileal crypt in WT mice and DR5^-/- mice (n = 6). At least 15 crypts were counted randomly for each slide and the mean value was calculated. F qRT-PCR analysis of Hmgcs2 and CDX2 in ileum of WT mice and DR5^-/- mice (n = 6). G The protein expression of CDX2 in ileum of WT mice and DR5^-/- mice (n = 4). H Representative images for immunohistochemical staining of CDX2 (scale bars: 100 μm) and quantification of CDX2⁺ cells in ileum of WT mice and DR5^-/- mice (n = 6). At least 15 correctly aligned villi including crypts were counted randomly for each slide and the mean value was calculated. I qRT-PCR was performed to examine the expression of ISC lineage differentiation genes in ileum of WT mice and DR5^-/- mice (n = 6). Elf3 for enterocyte fate specification, Klf4 for goblet cell fate specification, Ngn3 for enteroendocrine cell fate specification, Mmp7 and Sox9 for Paneth cells fate specification were detected in the ileum.