Mouse Embryonic Stem Cells Inhibit Murine Cytomegalovirus Infection through a Multi-Step Process (original) (raw)
Figure 3
Comparison of transfected/integrated MCMV IE promoter activity in MEF and ES cells.
(A) Results of the galactosidase enzyme assay on transgenic ES cells and MEF. β–gal positive cells (blue). (B) β–gal activity in wild-type MEF and transgenic ES cells and MEF. Activity could only be detected in transgenic MEF, at 7.9 µU/µL. (P<0.001) (C, D) Detection of promoter activity by flow cytometric analysis of GFP reporter expression. (C) The percentage of GFP positive cells with MCMV IE promoter activity and EF-1α promoter activity. (D) MCMV IE promoter activity after normalization with EGFP expression values under the control of the EF-1α promoter. There was a significant difference between ES and NIH3T3 after normalization (P<0.05). (E, F) Examination of the influence of histone modification and stimulation of the cAMP pathway on the transfected MCMV IE promoter in ES cells. (E) Co-treatment with FSK and TSA drastically increased the number of GFP-positive cells. (F) FSK and TSA each increase GFP fluorescence activity when administered individually (FSK: P<0.01, TSA: P<0.01), but the strongest response was seen when they were administered together (P<0.001). All presented experiments were performed at least 3 times, and data are given as the mean±SD. * P<0.05, ** P<0.01, *** P<0.001, _t-_test.