Figure 2.

Isolation of glomeruli and real-time PCR for selected genes. (a) Glomeruli (g) were isolated by perfusion of magnetic beads (arrows) that accumulated in the glomerular vessels. (b) Isolated glomeruli retain intact morphology with the majority simply comprising the tuft (left panel) whereas on occasions the tuft was surrounded by Bowman's capsule (arrows, right panel); (c) RNA integrity was preserved. FU, fluorescence units. Real-time quantitative reverse transcriptase–PCRs (qPCRs) for (d) Acy3, (e) Cyp4a12a, (f) Treh, and (g) Hsd3b2; transcripts positively correlating with albuminuria in array analyses. qPCRs for (h) Pln and (i) Trf, transcripts negatively correlating with albuminuria in array analyses. F, female; M, male. Hypoxanthine-guanine phosphoribosyltransferase (hprt) was used as a housekeeping gene. Fold changes in expression are expressed relative to B6 female mice where average expression was given an arbitrary value of 1 (a=P<0.05, b=P<0.01, and c=P<0.001 compared with B6 female mice; d=P<0.01 compared with B6 male mice; e=P<0.001 compared with B6 male mice; f=P<0.001 compared with FVB/N female mice, n=4 in each group). Bar in a is 50 μm and in b is 20 μm. Data were log transformed before analysis and are presented as geometric means and confidence interval.