Fluorofenidone offers greater protection of glomerular expansion at earlier than later stages of DN pathogenesis.

<p>A) Experimental schedule. Six <i>db/db</i> or <i>db/m</i> mice were mock-treated with vehicle at the indicated time points; six <i>db/db</i> mice were given either fluorofenidone (FD) or losartan daily, starting at 5, 8, or 12-week old; all groups were treated until the age of 24-weeks old. Duration of treatments for all schedules is indicated. B) Typical PAS staining images of glomeruli for the respective animal groups indicated in panel A. Ctrl (control): mock treatment. Scale bars represent 20 µm. C) Glomerular expansion index was derived by quantification of glomerular expansion based on PAS staining; means±SD are graphed. *: <i>p</i><0.05 in comparison to <i>db/m</i>; #: <i>p</i><0.05 in comparison to the respective Ctrl (diabetic) mice; $: <i>p</i><0.05 in the indicated comparison. C: Ctrl, F: fluorofenidone. L: losartan. D) Urinary albumin excretion was determined using 24-hour urine collections (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111242#pone-0111242-t002" target="_blank">Table 2</a> for details). As comparable levels of UAE were obtained in 5, 8, and 12-week old mock-treated animals, the average UAE for mock-treated <i>db/db</i> mice as a single entity was calculated; this was used to derive the percentages of UAE in fluorofenidone and losartan-treated <i>db/db</i> mice. Six mice were used in individual groups.