FIG. 7.
Inhibition of ATF4 or suppression of ER stress ameliorates inflammatory gene expression and reduces vascular leakage in STZ-induced diabetic mice. A: Adult C57BL/6 J mice were given intravitreal injection of Ad-GFP or Ad-ATFWT (1 × 109 viral particles per eye). Two weeks after adenoviral injection, retinas were dissected and expression of proinflammatory factors (TNF-α and ICAM-1) was determined by Western blot analysis and quantified by densitometry (mean ± SD, n = 8). **P < 0.01. B: Four-week-old diabetic mice were randomly selected to receive an intravitreal injection of Ad-GFP or Ad-ATF4DN (1 × 109 viral particles per eye). Two weeks after injection, retinal levels of VEGF and ICAM-1 were evaluated by Western blot analysis (mean ± SD, n = 3). C and D: After onset of diabetes, STZ-induced diabetic mice received periocular injections of PBA (0.4 μmol/eye) into one eye and the same amount of vehicle into the contralateral eye twice a week for 6 weeks. Expression of VEGF in the retina (C) was measured by Western blot analysis and quantified by densitometry (mean ± SD, n = 4). Retinal vascular permeability (D) was measured by Evans blue albumin method. Results were expressed as microgram per milligram total retinal protein (mean ± SD, n = 6). Immunostaining of VEGF and GS in retinal sections from diabetic mice after PBA treatment (E) or Ad-ATF4DN treatment (F) as described above. Images represent four animals in each group. PBS, phosphate-buffered saline. (A high-quality digital representation of this figure is available in the online issue.)