Induction of EPO expression by Wt1(–KTS) in human embryonic kidney (HEK293) and hepatoma-derived HepG2 cells. (A, top row) Transcripts of EPO, Wt1, and β-actin were detected by reverse transcription PCR (amplification with 32 cycles) in human embryonic (HEK293) kidney cells stably transfected with Wt1(–KTS), Wt1(+KTS), or empty expression vector (pCB6+). (A, bottom row) Expression of EPO, Wt1, and β-actin in transiently transfected HepG2 cells. cDNA was diluted 1:10 with H2O for amplification of β-actin. RT-PCR products were run on 1.5% agarose gels and stained with ethidium bromide; for better visualization, gel photographs are presented as negatives of the originals. (B) Induction of EPO expression by Wt1(–KTS) was confirmed by immunoblotting of EPO protein in whole cell extracts of HEK293 cells. Wt1 immunoblotting reveals significant Wt1(–KTS) expression. α-Tubulin was detected as internal control of equal amounts of protein.
