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Phototoxic Action Spectrum on a Retinal Pigment Epithelium Model of Age-Related Macular Degeneration Exposed to Sunlight Normalized Conditions

Figure 4

Characterization of the autofluorescence in A2E-loaded RPE cells.

A. Absorbance of RPE cells treated with 40 µM A2E (A2E+RPE (a), solid line) or A2E-untreated (RPE (b), dashed line). The curve ((a)–(b), dot line) representing the difference of absorption spectra between A2E-loaded RPE cells (a) and A2E-untreated RPE cells (b), shows absorption peaks at 335 nm and 440 nm. B. Absorbance spectra of free A2E in pure ethanol (solid line) or in modified DMEM (dashed line). Spectra are similar in both media and A2E displays maxima of absorbance at 335 nm and 440 nm. C. Emission spectra of RPE cells treated with 40 µM of A2E (A2E+RPE (a), solid line) or untreated (RPE (b), dashed line) under a 440 nm excitation. The curve ((a)–(b), dot line) representing the difference between the emission spectra in A2E-loaded RPE cells (a) and A2E-untreated RPE cells (b) shows a peak at 620 nm. D. Emission spectra of free A2E in pure ethanol (solid line) or in modified DMEM (dashed line) with a 440 nm excitation. Spectra are similar in both media and A2E displays a maximum of emission at 640 nm. E. A2E contents quantified by UPLC in RPE cells after 6 hours of incubation in various A2E concentrations (0, 5, 15, 20, 30 and 40 µM) in culture medium. The A2E content in RPE cells increases in a linear way according to the incubated A2E concentration. (n = 4, r2 = 0.9955). (RFU: Relative fluorescence unit).

Figure 4

doi: https://doi.org/10.1371/journal.pone.0071398.g004