FIG. 3.

Effects of tetracycline-mediated EWS/ETS expression on the expression and distribution of CD99 in UET-13 cells. (A) Relative CD99 levels in UET-13 transfectants in the absence or presence of tetracycline (Tet). UET-13 transfectants were treated with or without 3 μg/ml of tetracycline for the indicated periods. Real-time RT-PCR was performed to investigate the expression pattern of CD99. Signal intensities of CD99 were normalized using those of a control housekeeping gene (human GAPDH gene). Data are relative values with standard deviations from triplicate wells and are normalized to the mRNA level at 0 h, which is arbitrarily set to 1 in the graphical presentation. (B and C) Immunocytostaining of CD99 in UET-13 transfectants. Cells were cultured on coverslips in the absence or presence of tetracycline for 72 h and then stained with anti-CD99 antibody 12E7 (B) or O13 (C) as described in Materials and Methods. RD-ES cells were also examined as a positive control. For the staining of nuclei, DAPI was used.