Fig. 1

FGF2 induced paradoxical growth effects through p21 modulation in FGFR1 amplified and non-amplified ER + cells. A. Spheroid images of FGFR1 amplified cell lines (CAMA1 and MDA-MB-134) and non-amplified cell lines (MCF7 and T47D) show the effects of FGF2 simulation with various doses (5, 25, 125 ng/mL) for 14 days. Bar equals 1000 μm. B. Cell number estimation for images of spheroids in Panel A, with DMSO treatments serving as controls and set to a fold of one. Asterisks (*) indicate that the FGF2 treatment is significantly lower than the DMSO treatment (p < 0.05), while hashtags (#) indicate that the FGF2 treatment is significantly higher than the DMSO treatment (p < 0.05). C. Comparison of the levels of FGFR1 (full length and ICD) and p21 in ER + cell 3D cultures that are either amplified or non-amplified in FGFR1, with or without FGF2 treatment (25 ng/mL) for 72 h. The relative fold change in p21 for each cell line is determined by the ratio of p21 to β-actin in the FGF2 vs. control. D. Immunoblotting shows the expression of p21 in the 3D culture of ER + cells after exposure to various FGF2 dose treatments for 72 h. E. Immunoblotting shows p21 protein levels in 3D cell cultures treated with FGF2 (25ng/mL) plus p21 inhibitor UC2288 (2.0 and 5.0 µM) for 72 h. F. Spheroid images of all four cell lines show the effects of UC2288 (0.2 and 1.0µM) with or without FGF2 treatment (25 ng/mL) for 14 days. Bar equals 1000 μm. G. Relative fold change of ratio between cell number in + FGF2 vs. -FGF2 spheroids in each treatment for the images in Panel F, with the ratios in the DMSO treatment serving as controls and set to a fold of one