Figure 1
Endocytic AGEs uptake in cultured RAW264.7 cells. BSA, AGE-2, and AGE-3 were fluorescently labelled with Alexa Fluor 488. RAW264.7 cells seeded at 4 × 105 cells were incubated with fluorescent BSA, AGE-2, or AGE-3 at concentrations ranging from 0.2 to 200 μg/ml for the indicated periods: 10 min (a), 1 h (b), 2 h (c), or 4 h (d). Cellular uptakes of fluorescent BSA, AGE-2, and AGE-3 were determined by flow cytometry (n = 3). Each column represents the MFIs of Alexa Fluor 488-labelled AGEs relative to medium only (Unt, Untreated) after 10 min incubation, which was arbitrarily defined as 100%. Data are presented as the means ± SEM and were analysed using a one-way ANOVA followed by Tukey’s test. ***p < 0.001, **p < 0.01, *p < 0.05 compared with the value for Unt. The insert in (b) shows an overlay of a representative monoparametric histogram of CD204 expression in RAW264.7 cells incubated for 1 h with medium only (Untreated; greyscale), 200 μg/ml BSA (orange line), AGE-2 (blue line), or AGE-3 (red line).
