Figure 2

Patient-specific induced pluripotent stem cell clones were analyzed for pluripotency-associated gene expression. (A) Wilms tumor (WT)-induced pluripotent stem (iPS) cell, systemic lupus erythematosus (SLE)-iPS cell and autosomal-dominant polycystic kidney disease (ADPKD)-iPS cell clone expression of stemness-associated genes was analyzed by immunofluorescence and RT-PCR analysis. WT-iPS, SLE-iPS and ADPKD-iPS clones expressed high levels of OCT4, SOX2, KLF4 and NANOG. Immunofluorescence images were obtained at 40× magnification. (B) Total cellular RNA from WT-iPS, SLE-iPS and ADPKD-iPS clones was isolated and analyzed for endogenous pluripotency genes by RT-PCR. WT-iPS, SLE-iPS and ADPKD-iPS clones expressed gene transcripts of OCT4, SOX2, KLF4, NANOG, GDF3, hTERT and c-MYC. (C) For detection of silencing of exogenous pluripotency genes, BJ fibroblasts were infected with lentiviral vectors expressing OCT4, SOX2, KLF4 and c-MYC. Three days after viral vector transduction, RNA was isolated from infected fibroblasts, control uninfected fibroblasts, and WT-iPS, SLE-iPS and ADPKD-iPS clones. RT-PCR was performed for transgenes KLF4, c-MYC and OCT4. GAPDH gene transcript was amplified as an internal RNA control. No template (water) samples were included as controls.