Recombinant Histone and MMP-9 Expression

Core histones and MMP-9 proteins were expressed in Escherichia coli Rosetta 2 (DE3) pLysS cells (Novagen) and purified from inclusion bodies as described recently [20 (link)]. To generate mutant H3 and MMP-9 expression vectors, H3 and MMP-9 cDNAs were mutated by the QuikChange II site-directed mutagenesis kit (Agilent Technologies) before the construction. Further details of plasmid constructions are available upon request. G9a inhibitor BIX01294 is from Santa Cruz Biotech, and EZH1/2 inhibitor UNC1999 and MMP-9 Inhibitor I are from Sigma. Antibodies used in this study are as follows: H2A, H2B, H3, H4, and EZH2 antibodies from Abcam; H3K27me1 and EZH1 antibodies from Millipore; G9a, actin, and FLAG antibodies from Sigma; His antibody from Novagen; and MMP-9 antibody from Santa Cruz Biotech.

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Kim K., Shin Y., Kim J., Ulmer T.S, & An W. (2018). H3K27me1 is essential for MMP-9-dependent H3N-terminal tail proteolysis during osteoclastogenesis. Epigenetics & Chromatin, 11, 23.

Publication 2018

QuikChange II site-directed mutagenesis kit MMP-9 Inhibitor I MMP-9 antibody

Actin Antibodies Antibody Bix01294 Cdnas Cells Escherichia coli Ezh1 Ezh2 Histones Inclusion bodies Mmp 9 Mmp inhibitor i Plasmid Proteins Site directed mutagenesis Unc1999 Vectors

Corresponding Organization :

Other organizations : University of Southern California

Top 5 similar protocols

Variable analysis

independent variables

Mutant H3 and MMP-9 expression vectors

dependent variables

Expression of core histones and MMP-9 proteins

control variables

Wild-type H3 and MMP-9 expression vectors
G9a inhibitor BIX01294
EZH1/2 inhibitor UNC1999
MMP-9 Inhibitor I

positive controls

Wild-type H3 and MMP-9 expression vectors

negative controls

G9a inhibitor BIX01294
EZH1/2 inhibitor UNC1999
MMP-9 Inhibitor I

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