Figure 3From: Small molecule antagonist of the bone morphogenetic protein type I receptors suppresses growth and expression of Id1 and Id3 in lung cancer cells expressing Oct4 or nestinIsolation of lung cancer cells expressing Oct4 or nestin. Lung cancer cell lines were stably expressed with Oct4 or nestin promoters that drive GFP expression (Oct4/GFP and Nestin/GFP). (A) H1299 cell showing GFP expression after stable transfection with Oct4/GFP reporter. By FACS, GFP (+) cells were sorted from the Oct4/GFP cells and placed into cell culture. After 6 weeks cells were sorted for the GFP (+) and GFP (−) populations. (A) FACS analysis shows >99% purity of GFP (+) and GFP (−) cell populations. (B) Quantitative RT-PCR shows significantly higher expression of Oct4 and Sox2 in GFP (+) cells compared to GFP (−) cells (n = 3). (C) Immunoflourescent imaging showing Oct4/GFP (+) cells stain for Oct4. (D) The GFP (+) cells were sorted from H1299 cells stably expressing Nestin/GFP reporter and placed into cell culture. After approximately 8 weeks, the GFP (+) and GFP (−) cells were isolated by FACS. (E) Quantitative RT-PCR shows higher expression of nestin in GFP (+) cells compared to GFP (−) cells. (F) Dual immunoflourescent image shows NeuN (red) is not expressed in Nestin/GFP (+) (green) cells. (G-I) GFP (+) cells were isolated from Oct4/GFP and Nestin/GFP cells. Quantitative RT-PCR demonstrates differential expression of stem regulations and BMP type I receptors between Oct4/GFP and Nestin/GFP cells (n = 2).Back to article page