Figure 1

Expression and promoter activities of CD133 gene in human colon carcinoma Caco-2 and synovial sarcoma Fuji cell lines. A, Immunoblot analysis of CD133 protein in Caco-2 and Fuji cell lines. Human kidney cell line 293T and 293T transfected with pCR3.1-Uni-CD133 (293T/C) was used as a negative and a positive control, respectively. Actin is an internal control. B, Semiquantitative RT-PCR analysis of CD133 mRNA in Caco-2 and Fuji cell lines. cDNA from 293T and 100 ng of pCR3.1-Uni-CD133 plasmid (pCD133) were used as a negative and a positive control, respectively. The number of PCR cycles was at 32 for CD133. GAPDH is an internal control. C, FACS analysis of Caco-2 and Fuji lines with CD133/2-PE antibody. Numbers indicate the percentage of CD133-positive cells in the total population. The bold line represents a staining with CD133-PE antibody, and the dotted line represents the isotype control antibody. D, Schematic representation of the position of five CD133 promoters (P1 to P5) and exon1s (A to E). Gray boxes represent five first exons starting from the transcriptional start positions. Exons 1C2, 1D4, and 1E5 were identified in our previous study [24]. E, Promoter activity of P1, P2, P3, P4, and P5 in Caco-2 and Fuji cell lines. Left panel, schematic representation of luciferase reporter plasmids containing five CD133 promoters. +1 indicates the transcription start position of each exon1s. Right panel, luciferase activities of Caco-2 and Fuji cells transfected with five indicated reporters. Data are means ± s.d. of values from three independent experiments.