Skip to main content
. 2015 Dec 21;5:18709. doi: 10.1038/srep18709

Figure 2. G9a up-regulates PCL3 expression to promote EZH2 recruitment and H3K27 methylation of E-cadherin promoter.

Figure 2

(A) The protein levels of the PRC2 components of PANC-1 (P) and PANC-1-R (PR) cells were compared by Western blotting. (B) PANC-1-R cells were infected with control or G9a shRNA-containing lentivirus. RT-PCR analysis was performed to investigate the expression of the PRC2 complex components (left panel). RT-qPCR analysis was done to investigate the expression of PCL3 in PANC-1-R cells transfected with control or G9a shRNA or overexpressed methyltransferase-dead G9a (DN-G9a) (right-top panel). Protein levels of E-cadherin, PCL3 and G9a were studied by Western blot analysis (right-bottom panel). (C) Detection of G9a, PCL3 and E-cadherin expression in control or G9a-overexpressing PANC-1 cells by using Western blot analysis. (D) ChIP-qPCR analysis of the binding of PCL3 to E-cadherin promoter in control and G9a depleted cells. *p < 0.05. (E) PANC-1-R cells were infected with control or PCL3 shRNA lentivirus. After 48 h, expression of various target proteins was determined by Western blot analysis with indicated antibodies. (F) PANC-1-R cells were transfected with control or G9a shRNA in combination with FLAG-tagged PCL3. The protein levels of G9a, PCL3 and E-cadherin were studied by Western blot analysis with indicated antibodies. (G) ChIP-qPCR analysis was performed to investigate the binding of G9a to human PCL3 promoter in control or G9a-depleted PANC-1-R cells. *p < 0.05. (H) The binding of SETD1A and MLL to human PCL3 promoter and the methylation status of PCL3 promoter were studied by ChIP-qPCR in control or G9a-depleted PANC-1-R cells. *p < 0.05.