Figure 1.
spr-5 Is an H3K4me2 Demethylase. (A) Alignment of the C. elegans LSD1/KDM1 orthologs with human LSD1/KDM1. The key catalytic residues (N660 and K661) are shown at their relative positions within the amine oxidase domain, and the locations of the mutations are shown beneath each gene.(B) ChIP of H3K4me2 at
hop-1 in wild-type (N2),
spr-5(
by101), and
spr-5;
amx-1 mutants. The fold enrichment (of the percentage of input precipitated) in H3K4me2 Ab over no Ab is shown, and the positions of the primer sets in the promoter and gene body are indicated above.(C) Western blots of wild-type (N2) and
spr-5;
amx-1 protein extracts probed with actin, histoneH3, and H3K4me2 (positions marked). The ratio of H3K4me2/histone H3 normalized to actin is shown below.(D-H) Immunofluorescence with (D and G) SPR-5 N and (E and F) C-terminal Abs showing (D) the adult gonad, (E) oocytes, (F) an early embryo, (G) sperm, and (H) the corresponding DAPI image of sperm. The insets in (E), (F), and (G) are zoomed in on part of the panels. The identical staining pattern was observed with both N- and C-terminal Abs.