Figure 2. .

Effect of diabetes on retinal Keap1 and its binding with Nrf2. (a) Gene expression of Keap1 was estimated in the retina by q-PCR and β-actin was used as housekeeping gene. (b) Protein expression of Keap1 was determined by Western blot technique using β-actin as loading control. Binding of Keap1 with Nrf2 was determined in the retinal (c) homogenate by precipitating Keap1 followed by Western blotting for Nrf2, and (d) in the cytosolic fraction by precipitating Nrf2 and Western blotting for Keap1. Values from normal rat retina are considered as 1 (for mRNA) or 100% for protein expression. Each experiment was performed in duplicate in five or more rats in each group, and the values are expressed as mean ± SD. *P < 0.05 compared with the values obtained from normal rats.