Osmolarity and Glucose Differentially Regulate Aldose Reductase Activity in Cultured Mouse Podocytes
Figure 5
Expression of AR protein (a, b) and mRNA (c, d) in podocytes cultured in experimental media for 6 hours and 5 days, as described in Materials and Methods. (a) 15 μg protein from cell lysates was immunoblotted using rabbit polyclonal anti-AR antibody and visualized with BCIP-NDT yielding ~37 kDa bands for AR and ~47 kDa bands for α-smooth muscle actin. (b) Results of densitometric quantification of corresponding immunoblot bands are expressed as optical density (OD) ratios of AR to actin bands. (c) 2 μg total RNA was reverse transcribed and resulting cDNAs were amplified in 26 cycles, using specific primers for AR and for β-actin (see Table 2). 18 μL of PCR products were electrophoresized in 1.4% agarose gel and stained with ethidium bromide. (d) Results of densitometric quantification of corresponding RT-PCR products are expressed as optical density (OD) ratios of AR to actin bands. * versus NG-Nosm, ** versus NG-Nosm and HG-Nosm 5-d, *** versus NG-Nosm and HG-Hosm 6 h. a.u., arbitrary units.
