Fig 2: Schema of conditional KO of the ENDRA gene. The Pax8 promoter drives expression of rtTA which requires doxycycline to activate the bicistronic Cre recombinase and luciferase-expressing transgene. Cre is expressed specifically in the nephron but not in glomeruli. ETA receptor KO mice are homozygous for the floxed ENDRA gene and heterozygous for the two transgenes.

Fig 3: Increased infiltration of F4/80 positive macrophages is associated with ETBR expression in smoking induced chronic inflammation: Three color immunofluorescence images of the ETAR (purple), ETBR (red) and F4/80 (green) expression in the pancreas of WT and KC mice that were either sham treated or exposed to cigarette smoke. The representative images are shown in Panel A where the areas of overlap are highlighted in a box and zoomed (inset). The degree of overlap of F4/80 staining with ETAR and ETBR was measured using ImageJ to calculate Manders overlap coefficient (B and C respectively). (* p < 0.05, ** p < 0.005, ns = not significant) (Scale bar = 20 µm; zoom scale bar = 10 µm).

Fig 4: Representative photomicrographs of kidneys from Pax8-rtTA/LC1/floxed ETA (iETA) mice before and after doxycycline (DOX) treatment. Panel A shows cortex and Panel B shows medulla prior to DOX treatment; Panel C shows cortex after DOX treatment. DOX-treated iETA medullary staining is not shown since ETA immunostaining was not apparent in untreated iETA mice. Arrows indicate blood vessels. Staining was performed on 6 different mice of each genotype. Images are 200X.

Fig 5: Immunohistochemistry of ETAR.Cardiac left ventricular tissue from control animals with untreated cardiac arrest (A), or after 180 min of untreated return of spontaneous circulation (B), after 180 min of mild therapeutic hypothermia (C) or 180 min after S-PBN infusion (D) were stained with anti-ETAR. The amount of ETAR-positive cardiac cells was higher in pigs treated with mild therapeutic hypothermia (C) and S-PBN (D), respectively, compared to controls (A) and untreated animals (B).