Protein Complex Interactor Analysis and Differential Activity of KDM3 Subfamily Members Towards H3K9 Methylation
Figure 1
Enzymatic activity of KDM3 subfamily members towards H3K9 methylation.
Individual KDM3 subfamily members were transiently overexpressed in U-2 OS cells. (A-M) DAPI staining indicating cell nuclei. (A'-M') Cellular expression of Avi-tagged KDM3 subfamily members, as detected by streptavidin-AlexaFluor-488 recognizing the biotinylated Avi-tag. (A’’-M’’) H3K9me1, -me2 or -me3 groups, respectively, as detected by antibody staining. White circles outline the transfected cells in the last panel of each series. Note that cells transfected with KDM3A and KDM3B (A, D, G and B, E, H) abolish H3K9me1 (A’’ and B’’) and -me2 (D’’ and E’’) but not -me3 (G’’ and H’’) staining. On the other hand, JMJD1C transfection (C, F, I) does not decrease H3K9me1 (C’’), -me2 (F’’) or -me3 (I’’) levels. The catalytic mutant versions of KDM3A(H1120A) (J, L) and KDM3B(H1560A) (K, M) neither reduce H3Kme1 (J’’, K’’) nor H3K9me2 (L’’, M’’) levels. N shows the summary of the enzymatic activity described above.