Figure 5.

Raldh2 is expressed at sites of RA-signaling activity 72 hours after IR-AKI. Unilateral IR-AKI was performed in wild-type BALB/c mice, and kidneys harvested at the indicated times after injury. (A–D) Localization of Raldh2 expression at 0, 24, 72 hours, and 7 days after IR-AKI, as indicated. Kidney sections stained with anti-Raldh2 antibody, detected using horse radish peroxidase/3,3ʹ-diaminobenzidine substrate, counterstained with hematoxylin. (E) Co-localization of Raldh2 (red) with F4/80-positive macrophages (green) in the OM 72 hours after IR-AKI. White scale bar, 50 µm; black bars, 100 µm. (F) Expression of Raldh2 mRNA in renal macrophages 72 hours after IR-AKI. qRT-PCR for Raldh2 mRNA relative to Gapdh control mRNA was performed on RNA extracted from renal macrophages isolated using magnetic beads coated with anti-CD11b antibodies: uninjured mice (n=3), and mice 3 days after injury (n=8). Results expressed as mean±SEM fold change relative to uninjured controls. Two-tailed t test, **P<0.01 versus uninjured controls.