Fig. 7: Uncoupling self-renewal from adhesion in FOXM1-overexpressing JEB derived keratinocytes.

a Calculation of cumulative cell doublings in JEB, LAMB3-corrected, YAP-transduced, FOXM1-transduced JEB cultures derived from one representative experiment. Number of cell doublings was calculated as described in “Methods”. b CFE of keratinocyte cultures at passage 5, 7, 9, and 15 initiated from the control JEB patient and after LAMB3, YAP, or FOXM1 gene addiction. The number of cells per dish plated in the CFE is indicated between brackets and colonies were stained with Rhodamine B 12 days later. c Clonal analysis of normal keratinocytes (WT), untransduced, LAMB3-corrected, and FOXM1-transduced clonogenic JEB (see “Methods”). The percentage of holoclones, meroclones, and paraclones is indicated in red, light blue, and gray columns, respectively (n = 157 clones analyzed). d Top: adhesion of confluent cultured epidermal sheets prepared from normal keratinocytes (WT), JEB, LAMB3-corrected, YAP-transduced, FOXM1-transduced JEB29 cultures. The mere transportation of the flask from the incubator to the hood caused the spontaneous detachment of JEB and JEB-YAP and JEB-FOXM1 cultures, while WT and LAMB3-corrected keratinocytes remained firmly attached to the substrate. Bottom: CFE performed on the above cultures. The CFE was plated at a density of 4000 cells per dish, and colonies were stained with Rhodamine B 12 days later.