Figure 7

Smad2 phosphorylation at both c terminus and linker region is differentially affected by zoledronic acid according to ER status of breast cancer cells. A + B Representative immunofluorescent images of pSmad2C (green) and dapi staining (blue) in MDA-MB-231 cells (A) and MCF7 cells (B) treated with medium alone (C) or ZOL (Z) for 48 hours. C. Quantification of nuclear localisation of pSmad2C in MDA-MB-231 and MCF7 cells treated for 48 hours with medium alone (con) or ZOL (50 μM). Data represents minimum 100 cells per group, Mann Whitney U test for significance, NS = not significant D. Ratio of total cellular quantity of total Smad2/3 to pSmad2/3 in MDA-MB-231 cells treated for 1 hour with medium alone, ZOL (50 μM) or conditioned medium (CM) from MDA-MB-231 cells previously treated with ZOL (50 μM) or medium (control). Data represents 3 replicates and 3 repeats, Mann Whitney U test for significance, NS = not significant, *p = <0.05 E + F Representative immunofluorescent images of pSmad2L (green) and dapi staining (blue) in MDA-MB-231 cells (E) and MCF7 cells (F) treated with medium alone (C) or ZOL (Z) for 48 hours. G. Quantification of nuclear localisation of pSmad2L in MDA-MB-231 and MCF7 cells treated for 48 hours with medium alone (con) or ZOL (50 μM). Data represents minimum 100 cells per group, Mann Whitney U test for significance, NS = not significant, ***p value <0.001. H. Representative western blots for cellular quantity of pSmad2L and gapdh in MDA-MB-231 cells treated with medium alone (con) or ZOL (50 μm).