FIGURE 4.
Timp-3S156C/S156C mice have reduced TIMP-3 activity and increased expression of VEGFR-2. Spleen (a and b upper panels) and retina/choroids (a and b lower panels) of Timp-3+/+ (WT), Timp-3S156C/S156C (KI), or Timp-3S156C/+ (HET) were subjected to reverse zymography (a) and zymography (b). Retina/choroid lysates were immunoprecipitated (Ip) with anti-VEGFR-2 antibodies (c) followed by Western blot analysis with anti VEGFR-2 antibodies (middle panel) or anti-phosphotyrosine antibody (top panel). Lysates were immunoblotted with antibodies to actin (bottom panel) to standardize for protein loading. d, cryosections from WT and T3S156/S156 mouse eyes were analyzed by immunofluorescence staining with anti-VEGFR2 antibody using Texas Red-conjugated goat anti-mouse secondary antibody. Red staining indicated localization of VEGFR-2. INL, inner nuclear layer; ONL, outer nuclear layer; CC, choriocapillaris.