Fig. 5

FGFR1 overexpression upregulated p21 to inhibit cell cycle progression. A. Immunoblotting shows the levels of FGFR1, p21, and stat1/stat3 activation in 2D cultures of FGFR1 amplified cells (CAMA1 and MDA-MB-134) and non-amplified cells (MCF7 and T47D) after transfection with pCMV-XL6-FGFR1 plasmid or an empty pCMV-XL6 vector (as controls). B. Cell cycle phase proportions were determined by PI staining-based cell cycle FACS analysis in all four cell lines cells with FGFR1 and empty vector transfection in 2D culture. A ‘#’ symbol indicates G1 phase in FGFR1 transfected cells significantly higher than in empty vector transfected cells; C. 3D spheroid images are shown for four cell lines with FGFR1 and empty vector transfection after 14 days 3D culture. Bar equals 1000 µm. D. Relative fold change of cell number in FGFR1 transfected vs. vector control spheroids for images in (C). The cells transfected with empty vector are used as controls and set as fold one. E. Time course of cell number change over 14 days culture. for FGFR1 and vector control transfected cell lines. D and F, a ‘*’ symbol indicates the cells transfected with FGFR1 significantly lower than the cells transfected with empty vector (p < 0.05). F. cancer cell growth rate for cells transfected with FGFR1 and vector control. For D and F, a ‘*’ symbol indicates the cells transfected with FGFR1 significantly lower than the cells transfected with empty vector (p < 0.05)