Fig. 1. Changes in the pattern of DLG1 expression during the cell cycle. (A) Immunofluorescence analysis of DLG1 expression was performed on HaCaT cells using the anti-DLG1 monoclonal antibody 2D11. Various localisations of the protein can be seen in the three examples, including at sites of cell contact, diffuse cytoplasmic, nuclear excluded and also localisation to the mitotic spindle at mitosis and to the midbody at cytokinesis. Expanded insets indicate the spindle (upper panel) and the midbody (lower panel). (B) The specificity of the staining patterns seen in A with the anti-DLG1 antibody was verified by immunofluorescence analysis of DLG1 expression in a HaCaT cell line stably transfected with an shRNA against DLG1 (left hand panel). Also shown is the DAPI staining (middle panel) for the shDLG1 cells to show the nuclei and for comparison DLG1 expression in a control cell line (right panel) stably transfected with empty vector. (C) Immunofluorescence analysis of DLG1 expression during different phases of the cell cycle following arrest with aphidicolin (G1, 0 hrs) and subsequent release through to cytokinesis (12.5 hours). (D) FACS analysis showing cell cycle progression following the aphidicolin block and release.