FIG 8.

Cell division ceases after overexpression of Ar-SETD4. MKN45 cells were transfected with the pEGFP-C1 (GFP) and pEGFP-C1-Ar-SETD4 (Ar-SETD4) plasmids and then fixed with 4% paraformaldehyde. (A) For the BrdU incorporation assay, cells were incubated in medium containing 12 μM BrdU for 30 h and then fixed with 4% paraformaldehyde. (B) Detection of immunofluorescent signals of Ki67. The arrows indicate cells with GFP or Ar-SETD4 overexpression. Green, signals of GFP and GFP–Ar-SETD4; red, BrdU and Ki67 detected by Alexa Fluor 594-conjugated secondary antibody; blue, nuclei counterstained with DAPI. Bars, 20 μm. (C) Western blot analysis of the cell proliferation marker Ki67, PCNA, and H3S10ph after Ar-SETD4 overexpression. GAPDH and H3 were used as loading controls.