Figure 7. Assessment of endocrine cell induction in enteroids from Neurog3^WT or Neurog3^null mice or human intestines from normal and NEUROG3^DN proband.

(A) Neurog3^WT or Neurog3^null enteroids transduced with either constitutively active NEUROG3^WT- or NEUROG3^NULL-expressing lentivirus and grown in culture for 7 days. (B and C) Western blot of enteroids cell extracts isolated from (B) Neurog3^WT or (C) Neurog3^null mice. Enteroids were untransduced or transduced with NEUROG3^WT or NEUROG3^NULL and assessed for proteins using anti-Neurog3, Chga and tubulin antibodies. (D) Western blot of enteroids cell extracts isolated from a normal human (WT) subject untransduced or transduced with NEUROG3^DN and assessed 7 days later for proteins. (E) Human enteroids from WT and NEUROG3^DN subjects were examined for specified proteins and (F) mRNA. (G and H) mRNA isolated from murine (G) Neurog3^NULL and (H) Neurog3^WT enteroids transduced with NEUROG3^WT or NEUROG3^NULL and WT enteroids transduced with NEUROG3^WT and assessed 7 days later. (I and J) mRNA isolated from WT human enteroids and transduced with NEUROG3^DN and assessed 7 days later. (F–J) *P < 0.05, **P < 0.01, ***P < 0.005, 1-way ANOVA and Dunnett’s multiple comparison. (G–J) *P < 0.05, **P < 0.01, ***P < 0.005, unpaired Student’s t test.