Fig. 3

Liposomes functionalized with NCL-recognizing F3 peptide specifically bind and internalize into NB cells in vitro. A Binding to NB cell lines. Cells were incubated for 30 min at 4 °C with NT-Rhoda and T-Rhoda. Rhoda fluorescence associated with NB cells was evaluated by FC. CTR: NB cells only. B Co-localization of NCL and T-Rhoda. IMR-32 and SH-SY5Y cells were co-incubated with anti-NCL-A488 and T-Rhoda and analyzed by Imaging Flow Cytometry (IFC). BF: Bright Field; NCL: anti-NCL AlexaFluor488 moAb (green); 2nd Ab: AlexaFluor 488-conjugated secondary moAb; T-Rhoda: F3-nanocarrier (Red); Merge: co-localization (yellow). C Uptake by NB cell lines. Cells were incubated for 1 h at 37 °C with NT-Rhoda and T-Rhoda. Rhoda fluorescence associated with NB cells was evaluated by FC. Results are expressed as MRFI normalized over control cells. Columns: MRFI ± S.D. ***, p < 0.001: T-Rhoda vs NT-Rhoda. D Specific internalization into NB cell lines. SH-SY5Y and HTLA-230 cells were incubated as reported in C. After washing, samples were fixed and incubated with an anti-NCAM monoclonal antibody for plasma membrane localization. Green: N-CAM; Red: nanocarriers-Rhoda; Blue: DAPI. Bar: 30 μm