Supporting Information for Peptide Conjugation: Before or After Nanoparticle Formation? Sabrina Valetti,§,#,ɸ Simona Mura,§,# Magali Noiray,§,# Silvia Arpicco,ɸ Franco Dosio,ɸ Juliette Vergnaud,§,# Didier Desmaële,§,# Barbara Stella,ɸ,* Patrick Couvreur§,# § Université Paris-Sud, Faculté de Pharmacie, 5 rue Jean-Baptiste Clément, 92296 Châtenay- Malabry cedex, France # CNRS UMR 8612, Institut Galien Paris-Sud, 5 rue Jean-Baptiste Clément, 92296 Châtenay- Malabry cedex, France ɸ Dipartimento di Scienza e Tecnologia del Farmaco, Università di Torino, 9 via Pietro Giuria, 10125 Torino, Italy *To whom correspondence should be addressed. E-mail: [email protected] Tel.: +39 0116706800 Fax: +39 0116706663 1 TABLE OF CONTENTS Figure S1 3 Figure S2 4 Figure S3 5 Figure S4 6 2 Figure S1. Synthesis of the (5) squalene-peptide conjugate SQCKAAKN. 3 Figure S2. Western blot analysis. Representative Western Blot showing FZD-5 and β-actin expression in different pancreatic cancer and fibroblastic cell lines. MCF-7 cell line was used as a positive control. 4 Figure S3. Immunocytochemistry analysis and FZD-5 expression in MIA PaCa-2 cell line. Confocal images showing FZD-5 expression in MIA PaCa-2 cells. Nuclei were stained with Syto Red Fluorescent (blue). (a) Cells incubated with primary antibody against FZD-5 and Alexa Fuor®555conjugated secondary antibody (red). (b) Cells incubated with Alexa Fuor®555-conjugated secondary antibody to detect non-specific interactions (red). Scale bars: 20 µm. 5 Figure S4. Stability of N4-P*. Stability of N4-P* after 72 h incubation in water and cell culture medium (DMEM supplemented with 10% FBS) at 37 °C. Values represent mean ± SD. 6