Fig. 3. The Dsg3^dim population of adult palm keratinocytes shows a higher CFE than the Dsg3^bri population. (A) Dot plot showing the FSC and SSC of a suspension of cultured keratinocytes (a). Each dot represents one cell, and both FSC and SSC are measured in arbitrary units on a linear scale. Cells were sorted into two groups: fluorescence dim (Dsg3^dim: 20% of the cells with the least Dsg3) and fluorescence bright (Dsg3^bri: 40% of the cells with the highest Dsg3) populations (b). Alexa Fluor 488-conjugated IgG was used as the negative control (unfilled). (B) The sorted cells were plated onto the J2 feeder cells at 400 cells per well, and grown for 11 days in culture. Colonies were stained for keratins and visualised with anti-mouse HRP. More colonies grew in the Dsg3^dim population (a) than in the Dsg3^bri population (b). (C) Colony density analysis, as indicated by total colony area per dish (i.e. percentage of total area of dish) between Dsg3^dim and Dsg3^bri populations (mean±s.e.m. of triplicate samples). There is a higher total colony area in the Dsg3^dim, than in the Dsg3^bri, population and by 11 days the total colony area per dish in the Dsg3^dim population is significantly larger (^*P<0.05), nearly twofold that of Dsg3^bri. (D) CFE (ratio of colony number to plated cell number) at 11 days (mean±s.e.m. of triplicate). A slightly higher CFE is seen in Dsg3^dim than in the Dsg3^bri population. (E) Mean colony area (size) of Dsg3^dim and Dsg3^bri populations at three time points. The colony area increases with time and by 11 days the mean colony area in both Dsg3^dim and Dsg3^bri is significantly larger than that at 7 days or 9 days (^*P<0.05). In particular that of Dsg3^dim is twofold that of Dsg3^bri by 11 days (average 1.4 mm² vs 0.6 mm², ^**P<0.05).