Real-Time Monitoring of Apoptosis by Caspase-3-Like Protease Induced FRET Reduction Triggered by Amyloid Aggregation
Figure 1
Determination
of assay buffer and investigation of pFRET2-DEVD expressing B-TC-6 cells.
(a) Solutions
(KRHG, HBSS, RPMI without FBS, RPMI, Dulbecco´s modified medium, Schneider´s
drosophila medium, and H2O) were
analysed for autofluorescence at wavelength 535 nm (blue columns) and 480 nm (red
columns). Data presented are mean values ±SEM .
(b) Fluorescence at 535 nm and 480 nm was measured for KRHG buffer (black columns),
B-TC-6 cells expressing pFRET2-DEVD (green columns), and untransfected B-TC-6 cells
(grey columns). Data presented are mean values ±SEM . Difference in fluorescence signal between B-TC-6
cells expressing pFRET2-DEVD and untransfected B-TC-6 was considered to be
significant in a two-tailed unpaired t-test.
(c) Confocal image of pFRET2-DEVD
expressing B-TC-6 cells and (d) untransfected
B-TC-6 cells. Argon 488 nm and HeNe 633 nm lasers were used at the same energy levels in
the two images. Green fluorescence indicates expression of pFRET2-DEVD and blue fluorescence nuclear staining TO-PRO-3. Scale bars represent 10 m.
