Figure 3. RMECs growing on AGE-FN show enhanced JC-1 mitochondrial permeability A-D: Cells grown on native FN (A) were gated to exclude cell debris in a forward scatter versus side-scatter dot plot. This dot plot is expressed in terms of both the red and green fluorescence intensity after JC-1 incubation (B). The histogram charts the green (C) and red (D). E-H: The same gating schemes were used for RMECs growing on AGE-FN. Typical traces are shown from RMECs on AGE-FN (100 µM MGO) and demonstrate that AGE-exposed cells showed shrinkage, indicating apoptosis (compare A with E). There is also a relative increase in mitochondrial permeability in AGE-exposed cells as indicated by the net decrease in the red:green fluorescence intensity ratio (E-H). I: Quantification of JC-1 fluorescence revealed that RMECs cultured on AGE-FN (modified by 10–100 µM MGO) showed a significant increase in mitochondrial permeability (red:green fluorescence intensity ratio) in comparison to cells grown on native FN. (n=3; *p<0.05; comparison between AGE-FN and native FN).