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Fig. 3
Meis Regulates Hoxa2 Expression
(A) Meis and Pbx ChIP-seq-binding profiles at the HoxA and HoxB clusters in the IBA and IIBA of E11.5 embryos. Red boxes highlight Meis binding at Hoxa2 and Hoxb2 in IIBA chromatin.
(B) Meis occupancy at Hoxa2 promoter in IBA and IIBA chromatin (mouse) by ChIP qPCR. Pou6f2 is a positive control and Itih4 is a negative control (unbound region). Values represent the average of duplicate samples, and error bars indicate the SEM.
(C) Luciferase activity driven by Hoxa2 proximal promoter in HEK293T cells alone or in combination with Hoxa2, Meis, and Pbx expression vectors. Values represent fold activation over basal promoter activity, and are presented as the average of at least two independent experiments, each performed in triplicate. Error bars represent the SEM.
(D) Sequence conservation of the Hoxa2 proximal promoter in vertebrates, generated by the ECR Browser (Ovcharenko et al., 2004).
(E) Meis binding at hoxa2b promoter in zebrafish embryos by ChIP qPCR. Hoxb1a is a positive control; the negative control is a genomic region 10 kb upstream of the hoxba cluster. Enrichment of hoxa2b and hoxb1a is significantly higher compared to the negative control regions (p < 0.005). Values represent the average of three independent experiments, and error bars indicate the SEM.
(F) Whole-mount ISH on control MM and Meis-morpholino-injected embryos, using hoxa2b and dlx2 probes. Hoxa2b is downregulated in the second arch (gray arrows); dlx2 labels the developing branchial arches.
Reprinted from Developmental Cell, 32, Amin, S., Donaldson, I.J., Zannino, D.A., Hensman, J., Rattray, M., Losa, M., Spitz, F., Ladam, F., Sagerström, C., Bobola, N., Hoxa2 selectively enhances Meis binding to change a branchial arch ground state, 265-77, Copyright (2015) with permission from Elsevier. Full text @ Dev. Cell