Analysis of suspension growth and multinuclearity of cells over-expressing full-length MHCK-B, MHCK-B-Δ-WD, or MHCK-Δ-N-WD. (A) Cell densities were determined for wild type AX2 cells (◆), or for cells over-expressing full-length MHCK-B (■), the MHCK-B-Δ-WD truncation (▲), or the MHCK-B-Δ-N-WD truncation (●). Cells were grown in suspension culture (HL5 medium, 175 rpm shaking, 25°C) and cell densities were determined on the days indicated on the x-axis. Each plotted point represents the average value from four separate experiments and the vertical lines are the standard errors of those means. (B) Epifluorescent images of DAPI stained cells from the indicated cell lines were taken after four days of growth in suspension using an Olympus IX70 microscope system and an UPlanFL 20× objective lens. (C) The number of nuclei/cell was determined for each of the cell lines after four days of growth in suspension. The number of nuclei per cell was determined for a total of 250 cells per cell line over two separate experiments.