Fig. 3

Role of galectin-1 in regulation of FGF12 secretion A Galectin-1 knock-down in U2OS-FGF12-GFP.myc cells. Western blotting analysis of cell lysates of U2OS-FGF12-GFP.myc cells treated with siRNA against galectin-1 and scramble siRNA as a control. The amount of detected galectin-1 was quantified using densitometry measurements with ImageLab Software (right panel). Mean values from three independent experiments ± SD are shown. Statistical analyses were performed with Student’s t-test (*p < 0.05; **p < 0.005 and ***p < 0.001). B Impact of galectin-1 on FGF12 secretion. U20S and U2OS-FGF12-GFP.myc cells after galectin-1 knock-down were serum-starved at 37°C for 24 h and, after medium exchange, incubated at 42°C for 2 h. The medium from above the cells was collected, centrifuged and incubated with anti-myc-tag magnetic beads. 50% of eluted samples were loaded onto SDS-PAGE gels and analyzed by western blotting. FGF12 protein was detected with anti-FGF13 antibody, recognizing all proteins belonging to FHF family. The amount of detected protein was quantified using densitometry measurements in ImageLab Software (right panel). Mean values ± SD from three independent experiments are shown. Statistical analyses were performed with Student’s t-test (*p < 0.05; **p < 0.005 and ***p < 0.001)