Figure 1

A: Induced expression of EGFP in HFCL cells transfected with Egr-EF after exposure to 5-FU. EGFP expression in HFCL/EF cells transfected with Egr-EF was used to evaluate activation of the Egr-1 promoter by 5-FU treatment. EGFP expression in HFCL/EF cells exposed to 1, 2, 50, 100, 200, 300 mmol/L 5-FU was significantly higher than that in HFCL/EF cells not exposed to 5-FU and the HFCL group, P < 0.01. However, at 400 mmol/L 5-FU, there was no difference between the 5-FU treated group and the untreated group. Data are reported as mean ± SEM: B: EGFP expression confirmed by fluorescence inverse microscopy. (1) HFCL/EF group × 400. (2) HFCL/EF + 5-FU group × 400. C: Effect of supernatant from HFCL/EF cells treated with 5-FU on CD34⁺ cells. (1) HFCL/EF + 5-FU, (2) HFCL/EF, (3) HFCL/F + 5-FU, (4) HFCL/pCI + 5-FU, (5) HFCL + 5-FU. D: FL expression by HFCL/EF cells after Egr-1 promoter activation with 5-FU. FL production by HFCL/EF cells exposed to 5-FU (100 mmol/L) was measured using ELISA assay. Significant increases in FL protein levels were detected 24, 48, 72, 96, 120 and 144 h after exposure to 5-FU (P < 0.01, versus FL protein level at 0 h). Data are reported as mean ± SEM. E: (1)RT-PCR analysis of FL mRNA levels in HFCL/EF cells after 5-FU treatment. 1. 1.DNA Marker 11, 2.HFCL/EF with 5-Fu treatment group (in vitro), 3.HFCL/F without 5-Fu treatment group(in vitro), 4.HFCL/EF without 5-Fu treatment group(in vitro), 5.HFCL with 5-Fu treatment group (in vitro), 6.HFCL/EF plus 5-Fu group (in vivo), 7.HFCL/EF plus NS group(in vivo), 8. HFCL plus 5-Fu group (in vivo), 9. HFCL plus NS group(in vivo), 10. HFCL/pCI with 5-Fu treatment group (in vitro). (2) Western blot analysis of FL protein levels in HFCL/EF cells after 5-FU treatment. 1. 1. HFCL/EF without 5-Fu treatment group (in vitro), 2. HFCL/EF with 5-Fu treatment group (in vitro), 3. HFCL/pCI with 5-Fu treatment group (in vitro), 4. HFCL/F with 5-Fu treatment group (in vitro), 5. HFCL with 5-Fu treatment group (in vitro), 6. HFCL/F without 5-Fu treatment group (in vitro), 7. HFCL/EF plus NS group(in vivo), 8. HFCL/EF plus 5-Fu group(in vivo), 9. HFCL plus 5-Fu group(in vivo), 10. HFCL plus NS group(in vivo), Lain 2. the consistent expression of GAPDH in various groups.