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. 2019 Nov 14;48(1):421–431. doi: 10.1093/nar/gkz1037

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© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — Retention of RbBP5-binding activity of Cfp1 is required for appropriate H3K4 methylation. (A) Western blot analysis was performed on cellular extracts isolated following incubation of HEK393 cells with dox-inducible sh-RNA targeting Cfp1. Cfp1, actin, histone H3 and H3K4me3 levels were detected with indicated antibodies. (B) Immunoblotting of cellular extracts following the transfection of dox treated HEK293 cells with sh-resistant constructs of Cfp1 wild-type or mutant. Protein levels were detected as in panel 1A. The corresponding residues in Chaetomium thermophilum (Ct) are also indicated. (C) Immunoblotting of cellular extracts from Saccharomyces cerevisiae strain in which Cps40 was either deleted or rescued with flag tagged constructs corresponding to full length Cfp1 wild type or mutants. The corresponding residues in Chaetomium thermophilum (Ct) are also indicted. Flag-Cfp1, SET1, histone H3 and H3K4 methylation levels were detected with indicated antibodies. Arrows indicate full-length Cfp1 (red), and Cps40 constructs containing an internal deletion (blue) or a truncation of its N-terminus (green), respectively. The experiments were performed using in a SET762 background.