Fig. 6.

Butyrate protects Phb1-deficient ileal enteroids from dysbiotic microbiota-induced death. A Schematic of the experimental protocol of ileal enteroid treatment. Enteroids were cultured from ileal crypts of Phb1^fl/fl, Phb1^iΔIEC, or Phb1^ΔPC mice for 7 days and treated for 12 h with vehicle or 200 μg/ml ileal luminal feces filtrate obtained from 20-week-old Phb1^fl/fl, Phb1^iΔIEC, or Phb1^ΔPC mice. Concurrent with microbiota treatment, enteroids were treated with vehicle or 0.5 mM SCFAs alone or in combination. B, C Enteroid death as measured by morphological change during treatment with B Phb1^fl/fl or Phb1^iΔIEC microbiota filtrate or C Phb1^fl/fl or Phb1^ΔPC microbiota filtrate. D Representative enteroid images. Bar = 100 µm. E Enteroid death by morphological change. Enteroids were treated with 1 µM Mithramycin A for 1 h prior to microbiota filtrate/butyrate treatment as in A. n = 4–6 for all treatments. MithA, Mithramycin A. Results are presented as individual mice ± SD. *P < 0.05, **P < 0.01, and ****P < 0.0001