Fig. 4. Inh2 relocalization occurs within minutes of altering cell density. HeLa cells growing at high density were trypsinized and replated at low density on fibronectin-coated coverslips. After 20 (A,B,C), 40 or 60 minutes (D,E,F), cells were fixed with 3% paraformaldehyde and examined by immunofluorescence microscopy using an antibody to Inh2. Panels A and D show the staining of endogenous Inh2 in green. Panels B and E show the DAPI staining of DNA in red. Panels C and F show the merged image of the Inh2 staining with DAPI staining in red with the overlay of the two stains yielding yellow. Cells were scored as having nuclear staining brighter than, equal to, or dimmer than cytoplasmic staining. The bottom graph shows the combined result of three independent experiments with more than 100 cells being tallied in each experiment. The dashed line shows the percentage of cells with predominant nuclear staining and the solid line shows the percentage of cells with predominant cytoplasmic staining.