Inactive DNMT3B Splice Variants Modulate De Novo DNA Methylation (original) (raw)
Figure 5
DNMT3B isoforms drive unique and distinct localization, DNA staining, and H3K9me3 patterns in human cells.
FLAG-tagged DNMT3B2, DNMT3B3, and DNMT3B4 were transiently transfected into human HEK293 cells. 24 to 36 hours later, cells were fixed and stained with anti-FLAG (Sigma) and anti-H3K9me3 (Abcam) antibodies. Transfections were repeated at least in triplicate and cells were photographed with a fluorescence microscope. Representative DNMT3B (Globular, Diffuse, or Speckled: red), DNA (Condensed or Diffuse: blue), and H3K9me3 (Speckled, Spotty, or Faint: green) staining are depicted. Percentages of each type of staining versus DNMT3B isoform expressed are indicated below each image. The total number of independent cells analyzed: DNMT3B2, n = 237; DNMT3B3, n = 203; DNMT3B4, n = 215; none, n = 356.