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. 2017 Feb 1;144(3):400–408. doi: 10.1242/dev.143958

Fig. 2.

Fig. 2.

Deletion of Eed results in downregulation of Shh in the developing urothelium. (A,C) qRT-PCR analysis of Shh, Ptch1, Ptch2 and Pax6 expression relative to Gapdh in whole bladders from E18.5 control (Ctr, Eedf/+;ShhGC/+) and Eed cKOShh embryos. Control transcript levels were normalized to 1.0 and transcript levels in Eed cKOShh samples are displayed as fold change relative to control. (B,D) Whole mount in situ hybridization of control (Ctr, Eedf/+;ShhGC/+) and Eed cKOShh mutants at E15.5 using RNA probes for Shh (B) and Pax6 (D). PG, preputial glands; UP, urethral plate; U, urothelium. (E,F) ChIP-qPCR of Pax6 locus using anti-Ezh2 (E) and anti-H3K4me3 (F) antibodies. Schematic in E indicates location of PCR oligos (1-3) and transcription start sites (TSS) of Pax6. Open box, exon; arrows, direction of transcription. Data in A,C,E,F represent mean±s.e.m. of n=3-6. Unpaired Student's t-test, *P<0.05.