Fig. 4. CASK expression in HaCaT cells following release from cell cycle blockers. (A,B) Serum-starved HaCaT cells were re-stimulated to enter the cell cycle by adding serum. (A) Cells were treated with 10 µM BRDU for 45 minutes prior to staining with rabbit polyclonal anti-CASK antibody (green channel) and mouse monoclonal anti-BrdU antibody (red channel) at 12 and 22 hours. (B) Cells were co-stained with anti-CASK antibody (ZYMED; green channel) and mouse monoclonal anti-H3-P antibody for mitotic cells (red channel) at 22 and 30 hours. (C) Cells synchronised with Aphidicolin were stained with a rabbit polyclonal anti-CASK antibody (ZYMED; green channel) and mouse monoclonal anti-PCNA antibody (red channel). Corresponding cell-cycle analysis is shown in the graph at right. (D) Cells were synchronised with nocodazole prior to staining with a rabbit polyclonal anti-CASK antibody (ZYMED; green channel) and a mouse monoclonal anti-H3 antibody (red channel). Corresponding cell cycle analysis is shown at right. Scale bars, 50 µm.