The histone variant macroH2A confers functional robustness to the intestinal stem cell compartment
Fig 1
MacroH2A expression within the intestinal epithelium.
(A) Analysis of intestinal jejunum crypt or villus tissue fractions for macroH2A variant mRNA levels compared to mouse liver. ΔΔCT method, values normalized to Actb, N = 3 per condition, mean ± SD. (B) MacroH2A isoform mRNA level analysis within Lgr5-eGFPhigh CBCs or Hopx-tdTomato+ reserve ISCs FACS-purified from Lgr5-eGFP-IRES-CreERT2 or Hopx-CreERT2 Rosa26R-LSL-tdTomato mice. ΔΔCT method, values normalized to Actb, N = 3 per condition, mean ± SD. (C) Western blot showing macroH2A1 isoform protein level within FACS-purified populations of CBCs (again, Lgr5-eGFPhigh from Lgr5-eGFP-IRES-CreERT2 mice) or reserve ISCs (Hopx-tdTomato+ from Hopx-CreERT2 Rosa26R-LSL-tdTomato mice). Entire protein lysate from 30,000 CBCs or 20,000 reserve ISCs loaded into each well of gel corresponding to indicated samples on blot. (D) Immunohistochemical straining of pan-macroH2A1 or macroH2A1.1 in macroH2A WT or macroH2A DKO proximal small intestine. 10x objective. Scale bars = 100μm. **p<0.005, ***p<0.0005, ns = not significant, Student’s t-test.