Figure 5: A: Western blot for VEGF protein and the beta actin control the lanes Lane1: Control, Lane 2: AGE (100 μg/ml) treated, Lane 3: 2.5 mM glycine co-treated with AGE (100 μg/ml), Lane 4: 2.5 mM Glutamic acid co-treated with AGE(100μg/ml). It was observed that glycine decreased the VEGF expression.
B: VEGF RTPCR.
Lane 1: Negative control Lane 2: control, Lane 3: AGE (100 μg/ml), Lane 4: Glycine 2.5 mM+AGE (100 μg/ml), Lane 5: Glutamic acid 2.5 mM+AGE (100μg/ml), Lane6: DNA Ladder 600 bp. GAPDH was also run as a house keeping gene for the same.
C: The densitometric representation of the RTPCR gel normalized to GAPDH.
D: Vascular endothelial growth factor protein expression by immunofuorescence. a: Control (untreated), b: AGE (100 μg/ml) treated, c: Glycine (2.5 mM)+AGE (100 μg/ml), d: Glutamic acid (2.5 mM)+AGE (100 μg/ ml). There was an increase in the VEGF expression with AGE treatment and the amino acids glycine and glutamic acid decreased the expression when co-treated with AGE.
E: Real time PCR for VEGF mRNA expression is represented as a bar graph keeping control (untreated) as 1.