Fig. 1
Conditional macroH2A1.1 knockout (KO) in mice. a Upper panel. Targeting construct containing the sequence encoding mouse macroH2A1 (H2AFY), a loxP-flanked neomycin (neo) cassette 3′ of exon 6b (included in macroH2A1.2), and a rox-flanked cassette 3′ of exon 6a (included in macroH2A1.1). Lower panel. Targeting construct upon Cre-mediated excision. b Left: phase-contrast of first passage mouse embryonic fibroblasts (MEFs) after 3 days in culture. Right: protein lysates from MEFs of macroH2A1.1fl/fl and macroH2A1.1−/− (KO) mice were immunoblotted with anti-macroH2A1.1 and macroH2A1.2 antibodies. Histone H2A was used as a loading control. c macroH2A1.1 and macroH2A1.2 mRNA levels in adult mice tissues. RNA was extracted from the lung, liver, kidney, seminal gland, intestine, testis, brain, skeletal muscle, heart, and spleen of 3-month-old macroH2A1.1fl/fl, macroH2A1.1fl/− (HET), and macroH2A1.1−/− (KO) mice, and analyzed by qRT-PCR using isoform-specific primers. N = 5 mice/group. **p < 0.01; ***p < 0.001 relative to macroH2A1.1fl/fl