Figure 2.

The expression of miR-494 is ubiquitous in mice tissues and induced by I/R injury in the kidneys. (A) miRNAs were reverse-transcribed using miR-494 and U6 RNA-specific primers, and real-time PCR was performed as described in Concise Methods. The relative expression of the mature miR-494 was normalized to U6 RNA. Data are given as means ± SEM of triplicates. (B) The localization of miR-494 in the mouse kidney after I/R injury was detected by in situ hybridization. Paraffin-fixed sections of mouse kidney were hybridized with the digoxigenin-labeled miR-494 probe, proximal and distal tubular marker (aquaporin 1 [AQP1]), and Henle's loop marker (sodium-potassium-chloride cotransporter isoform 2 [SLC12A1]), and nuclei staining was stained by Contrast green. Figures are representative of three experiments performed on different days. Scale bar, 100, 200, and 500 μm as indicated. (C and D) Time course of the expression levels of both miR-494 and ATF3 in the kidneys after I/R injury. *P<0.05 (n=3, 5, or 6 mice per group as shown in the diagram). N.S., no significant difference.