Figure 2. Relationship between the duration of closed eye CL wear, corneal HIF-1α and VEGF levels. A: Corneal RNA was extracted and RT–PCR was performed with HIF-1α, VEGF, and GAPDH specific primers. C: western blotting analysis was performed as described in Methods. Corneal HIF-1α and VEGF levels were normalized to that of GAPDH. B, D: Densitometry analysis expressed in arbitrary units as the mean±SD, n=3, **p<0.01, as compared with HIF-1α of normal cornea. △p<0.01, as compared with VEGF of normal cornea.