Figure 2
Confocal laser scanning immunofluorescence microscopy for AGE uptake. Prior to the experiments, RAW264.7 cells were fluorescently labelled using PKH26 (red) and seeded at 4 × 105 cells/well followed by culturing for 10 min and 4 h with 200 μg/ml BSA, AGE-2, or AGE-3, all of which were fluorescently labelled with Alexa Fluor 488 (green). The incorporated fluorescent BSA, AGE-2, and AGE-3 in cells was determined by confocal fluorescence microscopy. Each panel of BSA, AGE-2, or AGE-3 indicates, as follows; (upper) phase contrast, (upper middle) Alexa Fluor 488-labelled AGEs, (lower middle) PKH26-labelled cell membrane, or (lower) merge of Alexa Fluor 488 and PKH26. All micrographs in this figure were taken at the same magnification (×400). Scale bar indicates 50 μm. The clipped images at the upper left of the merge panels in AGE-2 or AGE-3 show higher magnification images of the white rectangle region in the corresponding panel. Scale bar indicates 10 μm.
