Reducing KDM5 affects cellular levels of oxidative stress.

<p>(A) Survival of wildtype control (<i>w¹¹¹⁸</i>) and <i>kdm5^10424/K06801</i> female larvae in 5% sucrose (left) or in 20 mM paraquat/5% sucrose (right) for six hours. * <i>p</i><0.01. (B) Western blot showing levels of KDM5 and the loading control histone H3 in wildtype (<i>w¹¹¹⁸</i>) and <i>kdm5^10424/K6801</i> adult female heads (three heads per lane). (C) Survival curve of <i>w¹¹¹⁸</i> and <i>kdm5^10424/K06801</i> adult females fed 20 mM paraquat in 5% sucrose. Adults were one to three days old at the start of the experiment. Error bars represent standard error. The two survival curves are significantly different from one another (<i>p</i><0.05). (D) Detection of oxidized proteins in wildtype and <i>kdm5^K6801</i> homozygous mutant 3^rd instar larvae using oxyblot. Molecular weight markers are shown on the left. – indicates negative control lanes in which carbonyl groups were not derivatized. <i>kdm5</i> mutants show ∼2-fold increase in total levels of oxidized proteins. Histone H3 is shown as a loading control. (E) Schematic of <i>lacZ</i> mutation reporter assay. Adapted from Garcia et al <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1004676#pgen.1004676-Garcia1" target="_blank">[51]</a>. (F) <i>lacZ</i> mutation frequency of wildtype control (<i>w¹¹¹⁸</i>) and <i>kdm5¹⁰⁴²⁴</i> homozygous mutant 3^rd instar larvae. Genotypes are <i>w¹¹¹⁸</i>; #9lacZ; + (labeled <i>wt</i>) and <i>w¹¹¹⁸</i>; <i>kdm5¹⁰⁴²⁴</i>, #9lacZ; + (labeled <i>kdm5^−/−</i>. * <i>p</i><0.01. (G) Generation of wildtype clones in a wildtype background in larval wing imaginal disc. Genotype is hs-FLP¹²²/+; FRT40A/FRT40A ubi-GFP. Areas circled with dashed line have two copies of ubi-GFP (twin spots). Areas shown with a solid line have no GFP. (H) Generation of <i>kdm5^K6801</i> mutant clones in wing imaginal disc. Genotype is hs-FLP¹²²/+; <i>kdm5^K6801</i> FRT40A/FRT40A ubi-GFP. Areas shown with a dashed line have two copies of GFP (twin spots). Areas circled with sold line are homozygous mutant for <i>kdm5^K6801</i>. (I) Quantitation of clone area in E (<i>wt</i>; N = 6) and F (<i>kdm5</i> mutant; N = 8). ns = not significantly different.