Figure 3.

CDK2 Phosphorylates MLL2 to Regulate Its Binding

(A) Cell-cycle Fucci profiles of untreated and CDK2 inhibitor (CDK2I, CVT-313, 20 μM) treated cells for 4 hr.

(B) qRT-PCR analysis of transcript levels in ESCs treated with CVT-313 for 4 hr. Data are the average of three independent replicates.

(C) qChIP for MLL2 and MENIN in ESCs treated with CVT-313 for 4 hr. Data are the average of three independent replicates.

(D) MLL2 immunoprecipitations (IPs) from ESC lysates (200 μg protein; ±CVT-313) were probed with MLL2, phospho-threonine (pTP), or phospho-serine (pSP) antibodies. IgG was used as an IP control, and whole cell lysate was immunoblotted (input) alongside IPs.

(E) ESCs were transfected with a construct expressing constitutively active (CA) or inactive (KE mutant) CDK2-CCND1 gene fusion driven by CAGi promoter, and then qChIP assays for MLL2 were performed 36 hr later for GATA6 and SOX17. Data are the average of three independent replicates.

(F) Model depicting the cell-cycle control of bivalent domains. All data are representative of biological replicates.

Error bars in this figure represent the SEM. ^∗p < 0.05.