Figure 6: Exogenous CnB uptake is MD2-dependent.
(a) Co-localization of CnB and MD2. The MD2-cherry-transfected Hek293 cells were co-incubated with CnB-GFP for 30 min, fixed, and visualized using a LSM700 confocal laser scanning microscope (63×, scale bar 10 μm). (b,c) Influence of MD2 knock down on CnB uptake. The influence of MD2 knock down on CnB uptake was analysed by western blot analysis (b) or FI (c). (d) Co-IP of CnB and MD2. The MD2-transfected HEK293 cells were lysed, and the lysates were co-incubated with CnB-GFP or GFP for 2 h at 4 °C. Next, rabbit anti-MD2 pAbs or rabbit IgG1 were added and incubated overnight. Protein A beads were used to capture the complex for 2 h, and the beads were then washed five times and boiled for 5 min. The interaction was detected by western blot analysis of anti-CnB antibody. Data represent mean ± s.e.m from two independent experiments. **P < 0.01 (t-test, two-tailed).
