FIGURE 7.

MEP50 and PRMT5 regulate differentiation and proliferation in an epidermal equivalent model. KERn were electroporated twice with control or MEP50 or PRMT5 siRNA and seeded for epidermal equivalent culture. After 4 days of exposure at the air-liquid interface, the equivalents were harvested and sectioned. A and B, MEP50 and PRMT5 are required for appropriate skin equivalent formation. KERn were electroporated twice with control, MEP50, or PRMT5 siRNA and seeded for epidermal equivalent culture. After 4 days of exposure at the air-liquid interface, the equivalents were harvested and stained with hematoxylin and eosin. The nylon support membrane is indicated by an asterisk, and the extent of the epidermis is indicated (E). Similar results were observed in three separate experiments. The graph compares epidermal equivalent thickness among control, MEP50, and PRMT5 siRNA cultures. The values are mean ± S.E. (n = 3). *, p < 0.005. Scale bars = 100 μm. C and D, MEP50 and PRMT5 are required for cell proliferation. KERn were electroporated twice with control, MEP50, or PRMT5 siRNA and seeded for epidermal equivalent culture. After 4 days, the equivalents were stained with Ki67 antibody and Hoechst. The membrane (asterisk) and extent of epidermis (E) are shown. The graph quantitates the number of Ki67-positive cells in the control, MEP50, and PRMT5 siRNA rafts. Similar results were observed in three separate experiments. Significant differences were determined using Student's t test (*, p < 0.005). Note that the blue-green spots are Ki67 staining compared with the larger stained circular structures, which are nonspecific staining to the nylon membrane.