Generation of HaCaT cell lines stably depleted of hDlg1 or hScrib expression. Panel A.

HaCaT cells were transfected with control plasmid (TR2) or plasmids with targeting sequences specific for either hDlg1 or hScrib. After selection, colonies were picked and levels of hDlg1 and hScrib expression were ascertained by western blotting. A selection of clones is shown at different passages, with the levels of tubulin expression shown for comparison. Panel B. Morphological appearance of the control (TR2 series) and representative hDlg1 and hScrib-depleted cell lines at low and high density. Note the typical HaCaT cobblestone morphology of the control cells and hDlg1-depleted cells, and the more fibroblastic appearance of cells in the hScrib-depleted clones at low cell densities. At higher cell density all three lines exhibit similar cobblestone morphology (right hand panels).