Fig. 4.
Inhibiting KDM5 restores altered H3K4 methylation levels upon BRWD3 depletion. (A) Violin plot of quantitative IF assays in Drosophila S2 cells using an anti-H3K4me3 antibody (A) or an anti-H3K4me1 antibody (B). Five micrograms of KDM5 dsRNA was used in codepletion. Each distribution represents the signal intensities of 1,000 randomly selected cells from three biological replicates. ****P < 0.0001 using the ANOVA one-way analysis with Tukey’s multiple comparisons test. (C) Loss of a single copy of BRWD3 gives a Su(var) phenotype that is dependent on KDM5. (D) A model for BRWD3-dependent regulation of H3K4 methylation levels. BRWD3 targets KDM5 for degradation. In the absence of BRWD3, KDM5 activity is increased resulting in demethylation of H3K4me3, which generates increased H3K4me1 levels.