Figure 3
Functional EpoR is expressed on murine and human germline-derived immortalized cell lines. Panel A. (a) RT-PCR analysis of EpoR expression in the P19 murine teratocarcinoma cell line. Upper panel, RQ-PCR data, lower panel, SDS-PAGE gel of RT-PCR amplicon. The experiment was performed twice with similar results. (b). The effect of erythropoietin (0–100 U/ml) on adhesion of P19 cells to fibronectin. Combined data from three independent experiments are shown. (c). The effect of erythropoietin (0–100 U/ml) on the chemoattraction of P19 cells across Transwell membranes covered with gelatin. Combined data from three independent experiments are shown. (d). Phosphorylation of AKT and p42/44 MAPK in P19 cells. A representative blot is shown. Panel B. (a) RT-PCR analysis of EpoR expression in the NTERA2 human teratocarcinoma cell line. Upper panel, RQ-PCR data; lower panel, SDS-PAGE gel of RT-PCR amplicon. Experiment was performed twice with similar results. (b). Expression of EpoR on the NTERA2 cell line as measured by FACS. One representative histogram out of three experiments is shown. (c). Effect of erythropoietin (0–10 U/ml) on adhesion of NTERA2 cells to fibronectin. Combined data from three independent experiments are shown. (d). Phosphorylation of AKT and p42/44 MAPK in NTERA2 cells. One representative blot out of two is shown. Panel C. (a) RT-PCR analysis of EpoR expression in the A2780 human ovarian cancer cell line (lane 1). Lane 2, positive control (NTERA2 teratocarcinoma cell line). Lane 3, negative control (H2O instead of cDNA). The experiment was repeated twice with similar results. (b). Effect of erythropoietin on chemoattraction of the A2780 ovarian cancer cell line across Transwell membranes. Combined data from two independent experiments are shown. (c). Phosphorylation of p42/44 MAPK in the A2780 human ovarian cancer cell line stimulated by erythropoietin. A representative blot is shown.