FIG. 1.

Isolation of RW-GFP cells, a subset of RWPE-1 cells with higher levels of s-SHIP promoter expression. (A) RT-PCR analysis of s-SHIP transcript in parental RWPE-1 cell line. (B) RWPE-1 cells were transfected with s-SHIP-GFP promoter reporter (11.5 kb-GFP) and few GFP⁺ cells were obtained (arrow, left panel), then isolated by FACS, and amplified in culture to create RW-GFP cells (right panels). (Scale bar=50 μm) (C) Flow cytometry analysis of green fluorescent protein (GFP) expression in RW-GFP cells (filled graph) after two rounds of cell sorting and cell amplification as compared with parental cells (empty graph). (D) Quantitative RT-PCR analysis of endogenous s-SHIP transcript expression in RW-GFP cells as compared with parental RWPE-1 cells. Data represent mean values±standard error of the mean of three independent experiments, P values was determined by Student's test ***P<0.001.