Figure 5

(a) Relative expression levels (2^−ΔΔCt) of the RNA-seq of CBX2.1 downstream genes. ANGPTL4, NTRK1, CYP19A1, DMRT1, EMX2, ESR2 and KISS1 were upregulated after CBX2.1 overexpression (Over Exp). CBX2.1 silencing assay (siCBX2.1) reduced significantly the genes: ANGPTL4, DMRT1, ESR2 and KISS1. NTRK1, CYP19A1 and EMX2 gene did not show any expression in response to siCBX2.1. All graphs are the average of three independent experiments, error bars represent SD from the mean (SEM), and values are expressed as relative to control =1; ***P < 0.001; **P < 0.01; *P < 0.05. non-significant differences are not indicated. (b) Effect of CBX2.2 on DamID downstream targets: AKRC1, TGFα, AMIGO2 and RSPO3. Gene expression levels showed a substantial downregulation after CBX2.2 overexpression (Ov. Exp). The silencing of CBX2.2 (si CBX2.2) significantly stimulated the expression of AMIGO2, RSPO3 and AKR1C1 genes compared to scrambled siRNA. TGBβ2, NTRK2, FZD5 and SOX4 genes were significantly upregulated by CBX2.2. In the siCBX2.2 samples, NTRK2, FZD5 and TGFβ2 expression levels were found to be negatively regulated. TGFα and SOX4 expressions showed no effect relative to the scrambled sample. All graphs are the average of three independent experiments, error bars represent SD from the mean (SEM), and values are expressed as relative to control =1; ***P < 0.001; **P < 0.01; *P < 0.05. non-significant differences are not indicated.