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Fig. 8. Analysis of macroH2A distribution in T-antigen-transformed MEFs after conditional mutation of Dnmt1. (A,B) Male MEF line F11 was transfected with empty virus (A) or cre virus (B) to delete exons 4 and 5 of the Dnmt1 gene that encodes the methyltransferase core of the enzyme. Cells were fixed and subjected to combined immunofluorescence for macroH2A and kinetochore proteins (CREST) to mark chromocenters. (C,D) An independently derived cell line (F12) was subjected to similar mock (C) and cre-virus (D) transfections. F12 yielded results similar to those observed for F11. (E,F) A female MEF cell line (X17) was also subjected to mock (E) or cre-induced (F) loss of Dnmt1 function. Loss of Dnmt1 function in X17 and a second female cell line (X14, not shown) failed to perturb the localization of maroH2A.





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