Figure 1.
Effect of tumor necrosis factor-α (TNFα) on granulocyte-macrophage colony-stimulating factor (GM-CSF) production in human MDA-MB-231 cells. MDA-MB-231 cells were cultured in 6-well plates at a concentration of 1 × 106 cells/well. Cells were treated with vehicle and TNFα (2 ng/mL), separately. After 24 h incubation, cells and supernatants were collected. (A) Total cellular RNA was isolated and GM-CSF mRNA expression was determined by real-time PCR. (B) Secreted GM-CSF in culture media was determined by ELISA. (C) MDA-MB-231 cells were treated with vehicle or TNFα for 24 h and then were stained with GM-CSF (red) and DAPI (blue). White arrows indicate typical stained cells. (D) GM-CSF fluorescence intensity is shown. The results obtained from three independent experiments are shown. All data are expressed as mean ± SEM (n ≥ 3). ** p < 0.01, **** p < 0.0001 versus vehicle.