Fig. 4

KDM2B transcriptionally suppressed the expression of MOB1. a, PANC-1, AsPC-1 and MiaPaca-2 cells were treated with de-methylating agent 5-aza at the concentration of 5 μM for 72 h. Protein levels of MOB1 were analyzed by western blot (β-actin as internal control). b, KDM2B was knocked down in PANC-1 and AsPC-1 cells using shRNAs, and the expression of MOB1, H3K27me3, H3K36me2 and H3K4me3 and CTGF were tested using western blot (β-actin as internal control). c, PANC-1 and AsPC-1 cells were transfected with shKDM2B or control vectors. Chromatins were isolated and the binding of KDM2B and H3K27me3 to the MOB1 promoter was determined using ChIP assay as described in Materials and Methods (W: water; 2B: KDM2B; H3: H3K27me3). d, MOB1 promoter reporter was constructed. PANC-1 and AsPC-1 cells were co-transfected with 0.2 μg of the MOB1 promoter luciferase constructs pLuc-MOB1 and shKDM2B or control vector. Promoter activity was examined using a dual luciferase assay kit