Fig. 2

Binding of S100A6 to MDM2 in vivo and in vitro. A Co-immunoprecipitation assay for binding between endogenous S100A6 and MDM2 in MCF-7 cells. Cell lysates from MCF-7 were incubated with anti-S100A6 antibody or IgG in the presence of 0.2 M CaCI₂, 0.2 M NaCl, or 4 mM EGTA. The immunoprecipitates were detected by western blot using an anti-MDM2 antibody. Data from three independent experiments were presented as mean ± SD, which were to compare the effects of CaCI₂ and EGTA on the combination degree of MDM2 and S100A6 in S100A6-transfected MCF-7 cells. **P < 0.01. B A schematic diagram showing different domains of MDM2 and its truncations. C GST pull-down assay for binding between GST-S100A6 and HA-tagged full-length MDM2 or its truncations. D A schematic diagram showing different domains of S100A6 and its truncations. E GST pull-down assay for binding between HA-MDM2 and GST-fused full-length S100A6 or its truncations. F Confocal analysis showing the subcellular localization of MDM2 and S100A6. Scale bar: 5 μm