Protein Kinase Cε, Which Is Linked to Ultraviolet Radiation-Induced Development of Squamous Cell Carcinomas, Stimulates Rapid Turnover of Adult Hair Follicle Stem Cells
Figure 1
Hair follicle stem cells and clonogenicity of epidermal keratinocytes isolated from PKCε overexpressing mice and wild-type littermates. (a) is showing the representative gating of epidermal stem cell population in a dot plot from untreated WT, TG224, and TG215 mice. In each dot plot, the upper right quadrant is representing the CD34+/α6-integrin+ (double positive HSCs) stem cell population. Each value in the histogram is an average of FACS analysis of triplicate samples from keratinocytes pooled from two mice. (b) represents the total frequency of CD34+/α6-integrin+ keratinocytes in untreated indicated mice. (c) and (d) Clonogenicity of epidermal keratinocytes. Briefly, the keratinocytes from 7-8 weeks old indicated that mice were harvested using SMEM harvesting medium. Irradiated 3T3 cells seeded at density 106 cells/dish to the 60 mm dishes a day before seeding keratinocytes. For feeder layer, irradiated 3T3 cells were cultured in EMEM medium with 10% FBS and 1% penicillin-streptomycin. Equal numbers of keratinocyte cells (3000 cells/dish) were seeded for each type of mice and cultured with William’s E media for 2 weeks. For counting and measurement of colonies, dishes were fixed with 10% formalin and stained with 0.5% rhodamine B. (c) Keratinocyte colonies. Shown are the representative dishes of adult keratinocyte colonies from PKCε TG mice and their WT littermates. (d) Quantitation of colonies. The colonies were counted and colony size measured by using vernier caliper (Figure 1(c)). Each value is the mean ±SE of colonies from 4–7 dishes.
