Figure 1.

The expression of H19 is increased in cells, humans, and mice under ischemic/hypoxic conditions. H19 expression was determined in kidney transplant patients with short compared with long CIT (A) (n=5 patients per group), in mouse embryonic (E13.5), adult, and I/R kidneys (B) (n=8 mice per group). FACS-based sorting of mouse kidney homogenates after I/R injury exposed to rat anti–mouse-CD31-PE/rat anti–mouse-CD45-PerCP-Cy5.5 antibody for endothelial cells (C), and injured or healthy proximal tubular cells positive for lotus tetragonolobus lectin for proximal epithelium and anti-mouse Tim1-biotin antibody followed by streptavidin-APC (D and E). RNAscope analysis of H19 expression in control mice and mice after I/R injury and 24 hours of reperfusion (F), and 7 days of reperfusion (G) (n=5 mice each). Arrows indicate endothelial expression, asterisk indicates tubular epithelial induction. Expression of H19 in cultured endothelial cells (HUVECs) (H) and proximal tubular cells (RPTECs) (I), under hypoxic condition over a 72-hour time course. H19 expression after 16 hours of serum starvation or ATP depletion (n=4) (J). Each bar represents the mean±SEM of duplicate cultures, and the data are from a representative experiment of six independent experiments (unless otherwise stated) that were performed. *P<0.05; **P<0.01; ***P<0.001. A/DG, actinomycin A/2-deoxyglucose; NT, no treatment; TPL, kidney transplantation.