Figure 6.
ROS inhibit histone demethylation.
(A) Representative western blot for histone methylation markers and H4 protein loading control from nuclear extracts of UOK262 cells expressing cv and FH-FLAG plasmids. (B) Densitometric quantification of histone methylation markers relative to H4 loading control of UOK262 cv and FH-FLAG cells. (C) Measurement of demethylase activity of JMJD2A in vitro in the presence of fumarate, succinate, and N-oxalylglycine (NOG). (D) Measurement of demethylase activity of JMJD2A in vitro in the presence H2O2. (E) Measurement of synergistic inhibition of JMJD2A in vitro in the presence or absence of 1 mM fumarate and various doses of H2O2. Values are relative to activity of mock or fumarate treated cells without H2O2. In (B–E) the values denote mean + s.e.m. n = 3. *P < 0.05; **P < 0.01.