The histone variant macroH2A confers functional robustness to the intestinal stem cell compartment
Fig 6
MacroH2A’s influence of intestinal tumorigenesis.
(A) MacroH2A mRNA level analysis of healthy human intestinal crypt epithelium and human CRC cell lines. ΔΔCT method, values normalized to GAPD. N = 3 per condition, mean ± SD. (B) Graphical depiction of the H2AFY gene and its exons, including the mutually-exclusive exons of the macroH2A1.1 and macroH2A1.2 splice variants. (C) MacroH2A siRNA knockdown validation in RKO CRC cell line. ΔΔCT method, values normalized to GAPD independently per macroH2A primer relative to luciferace knockdown control. N = 3 per condition, mean ± SD. (D) MTT cell proliferation assay of RKO cell line during macroH2A1.1, 1.2, H2AFY, or control luciferace RNAi knockdown. N = 3 per condition, mean ± SD. (E) Left: representative H&E images of macroH2A WT and DKO Apcmin-derived tumors within the small intestine. 4x objective. Right: quantitation of average total tumors within entire small intestine of macroH2A WT and DKO. N = 8 mice per condition, mean ± SD. *p<0.05, **p<0.005, ***p<0.0005, ns = not significant, Student’s t-test. Scale bar = 100μm.