Rapamycin has little effect on the fibrogenic phenotype of tubular epithelial cells.

<p>HK2 cells or Animals were treated as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033626#s2" target="_blank">Methods and Materials</a>. A–C. Representative immunofluorescent costaining images of kidney sections derived from UUO mice or IRI mice, using anti-Kim-1 and anti-αSMA (A), anti-Kim-1 and anti-CD11b (B), anti-pS6K and anti-Kim-1 (C). Nuclei were labeled with DAPI (blue). Scale bar: 50 µm. D. HK2 cells were cultured for 48 hours in the absence or presence of aristolochic acid (AA), with or without administration of rapamycin. The cells were stained for Ki67 or pS6K. DAPI was used to stain the nuclei. E. Representative western blot and densitometric analyses for expression of CTGF, Collagen-I, and pS6K in cultured HK2 cells. β-actin was used in this experiment for equal protein loading control. Data were presented as mean ± SE. ** P<0.01, NS no significance. F. Assessment of proinflammatory and profibrogenic gene expression in culture HK2 cells. mRNA level of MCP-1, TGF-β, CTGF and Collagen-I were determined by realtime-PCR. ** P<0.01, NS no significance. Error bars represent S.E.