Figure 4.

Histone Methyltransferase Ash1l Is a Target of miR-291 and Regulates Hox Gene Expression

(A) Relative expression of Ash1l in DCR^fl/fl and DCR^−/− ESCs ± miR-291.

(B) Normalized luciferase levels after transfection of DCR^−/− ESCs with indicated vectors ± miR-291. Values were normalized to the no miRNA control. Bars represent mean ± SD of three independent experiments.

(C) ChIP-qPCR analysis of ASH1L enrichment at indicated loci. Data are represented as fold change of DCR^−/− over DCR^fl/fl signal.

(D) Relative expression of Hoxa10, Hoxd9, and Ash1l in DCR^−/− ESCs after transfection with the indicated miRNA or siRNA. Bars represent mean ± SD of three independent experiments. Data are represented as fold change compared to DCR^−/− ESCs.

(E) Western blot showing the levels of ASH1L in DCR^fl/fl and DCR^−/− ESCs mock transfected or transfected with miR-291 or siRNAs targeting Ash1l (siAsh1l). Numbers below each lane indicate the signal of each band normalized to the signal from HSP90. The normalized signal for DCR^fl/fl is set to 1.

(F) Schematic model of miR-290 regulation of PcG targeting and Ash1l.

See also Figure S4.