PMC

RAW264.7 cells were incubated with 30- and 150-nm gold colloids with or without the presence of: (A) an inhibitor of actin polymerization (cytochalasin D) or (B) an inhibitor of macropinocytosis (5(N,N,-dimethyl)amiloride hydrochloride). Each sample was analyzed in duplicate and the experiment was repeated twice. Each data point contained at least 100 multifocal images. Shown is mean value (n = 2).
*p < 0.05.

RAW264.7 cells were incubated with 30- and 150-nm gold colloids with or without the presence of inhibitors that block scavenger receptor and caveolin-mediated pathway. (A) Caveolin-mediated uptake was inhibited with filipin III. (B) Inhibition of both caveolin- and scavenger receptor-mediated uptake was achieved by incubating nanoparticles with siRNA bearing cells in the presence of filipin III. Each sample was analyzed in duplicate and the experiment was repeated twice. Each data point contained at least 100 multifocal images. Shown is the mean value (n = 2).
*p < 0.05.

RAW264.7 cells were incubated with 30- and 150-nm gold colloids with or without the presence of inhibitors that block the scavenger receptor and clathrin-mediated pathway. (A) Scavenger receptor was inhibited by MSR-1-specific neutralizing antibodies. (B) Expression of scavenger receptor was inhibited by siRNA. (C) Clathrin-mediated uptake was inhibited with chloropromazine hydrochloride. (D) Inhibition of both clathrin and scavenger receptor-mediated uptake was achieved by incubating nanoparticles with siRNA-bearing cells in the presence of chloropromazine hydrochloride. Each sample was analyzed in duplicate and the experiment was repeated twice. Each data point contained at least 100 multifocal images. Shown is mean value (n = 2). (E) Analysis of the SR-A expression by parental RAW264.7 cells and cells transduced with SR-A-specific siRNA bearing construct. Cell lysates were analyzed by western blot using monoclonal antibody directed against SR-A to control SR-A expression and anti-β-actin antibody to control protein loading.
*p < 0.05 when compared with negative control; **p < 0.05 when compared with isotype control.
SR-A: Scavenger receptor A.

Human sodium–citrate stabilized plasma obtained from three healthy donor volunteers was treated with NC, PC or nanoparticles as described in Materials and methods. Concentration of gold colloids was 50 μg/ml based on total amount of gold determined by the inductively coupled plasma mass spectroscopy. Each sample was analyzed in duplicate. Shown is a mean result (n = 2%CV <10). Cobra venom factor, used as a positive control, activates complement system through an alternative pathway, this is why no activation is seen in the C4d assay. The test results for the positive control in iC3b assay was high (1375.0 ± 0.05 pg/ml). The scale was changed to magnify the lower area; the arrow and the number above the PC value in the iC3b assay are used to show the actual response in this sample.
NC: Negative control; PC: Positive control.