Fig. 2

EZH2 enhance ISL1 expression through binding on Fragment 2. A) schematic reppresentation of the region tested for ChIP-qPCR in ISL1 (red segments) with distance relative to the 5? of Fragment 2 in kb. B) ChIP-qPCR fold change representing EZH2 bound genomic DNA relative to mouse IgG bound DNA. Androgen receptor (ar) genomic DNA as positive control6 and actin b as negative control. All fold change except for actin b are significant over IgG control. C) Western blot against EZH2 shows a succefull siRNA knockdown of EZH2 compared to control. D) ISL1 qPCR with EZH2 siRNA dispalys a reduced ISL1 signal. E) Western blot on EZH2 with control vector and EZH2 overexpression plasmid confirms the efficency of the overexpression. F) Luciferase assay with EZH2 overexpression (red bars) and control (gray bars). As before, vector (pGL3) and Fragment 2 forward show only little expression in both cases while EZH2 overexpression enchances the reporter activitiy of the plasmid with flipped Fragment 2 significantly.