Characterization of Pyrogenic Silica Nanoparticles

Pyrogenic amorphous silica nanoparticles (ASNP) NM-203 were from the same batch used in [19 (link)] and were obtained from the JRC Nanomaterials Repository (Ispra, Varese, Italy). The physico-chemical characterization of NM-203 ASNPs, before and after their suspension in different media, has been extensively reported elsewhere [17 (link)]. A/B Human serum and E. coli LPS (strain O55:B5) were from Sigma-Aldrich (Merck, Milan, Italy), while RPMI 1640 culture medium was provided by Corning (New York, NY, USA). The antibodies used were mouse monoclonal anti-GS (BD, Franklin Lakes, NJ, USA), rabbit polyclonal anti-LC3BI/II, rabbit polyclonal anti-p-NFκB, mouse monoclonal anti-NFκB, mouse monoclonal anti-LAMP1 (Cell Signaling, Danvers, MA, USA), rabbit polyclonal anti-TLR4 (Novus Biologicals, Cambridge, UK). All antibodies were diluted 1:1000 unless otherwise specified.