Figure 2

Ectopic JMJD3 promoted SAHF formation in WI38 cells. (a) Western blotting confirmation of the ectopic expression of JMJD3 and JMJD3H1390A after retroviral infection in WI38 cells. (b) WI38 cells were infected with JMJD3 and JMJD3H1390A for 6 days and then stained with SA-β-gal. Cells positive of SA-β-gal activity were calculated (Right). (c) WI38 cells infected with either JMJD3 or JMJD3H1390A were stained with 4,6-diamidino-2-phenylindole (DAPI), and the percentage of cells with DAPI foci was calculated. (d) WI38 cells from panel (c) were pulse-labeled with 5′-BrdU (bromodeoxyuridine) for 24 h and then immunofluorescence stained to detect the 5′-BrdU. A total of 100 cells were scored for 5′-BrdU incorporation. (e) Confocal fluorescence microscopy of SAHF markers (H3K9me3 and HMGA1) in WI38 cells and WI38 cells infected JMJD3 or JMJD3H1390A. Scale bars: 5 μm. Error bars represent the mean S.D. of triplicate experiments