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. 2017 Dec 9;8(2):491–503. doi: 10.1534/g3.117.300311

Figure 2.

Figure 2

β-catenin activity is not required for the expression of PRC2 components and bulk H3K27me3 levels. (A) FPKM values obtained from RNA sequencing assays in CM+ectoderm from En1Cre/+;R26R/+;β-cateninfl/∆ mutant and En1Cre/+;R26R/+;β-cateninfl/+ controls. (B) RT-qPCR analysis of the CM+ectoderm in En1Cre/+;R26R/+;β-cateninfl/+ (n = 7) and En1Cre/+;R26R/+;β-cateninfl/∆ mutants (n = 9). The data are represented as fold change in mutants over controls. The dotted line represents the β-catenin controls. Axin2 and Sox9 are known targets regulated by β-catenin. (C and D) Western blot analysis (n = 5) of EZH2 and H3K27me3 in CM+ectoderm from En1Cre/+;R26R/+;β-cateninfl/∆ mutants and En1Cre/+;R26R/+;β-cateninfl/+ controls. Band intensities were quantified using ImageJ. (E and F) Schematics representing the supraorbital CM in coronal sections near the frontal bone primordia. (G) Indirect immunofluorescence of SOX9, H3K27me3, and DAPI in the supraorbital mesenchyme (n = 2 controls; 3 mutants). Images were taken near the frontal bone primordia (plane I). Dashed lines indicate the brain and ectoderm boundaries. (*) indicates region of ectopic cartilage. Bar, 200 µm. CM, cranial mesenchyme; DAPI, 4’,6-diamidino-2-phenylindole; FPKM, fragments per kilobase of transcript per million mapped reads; n.s., not significant; RT-qPCR, reverse transcriptase-quantitative polymerase chain reaction.