Calpeptin treatment decreases cell density and proliferation.

<p>NS-TGFP cells were cultured in permissive conditions and treated with either 50 µM calpeptin or DMSO (control), and BrdU was added 3 h later as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033468#s4" target="_blank"><i>Materials and Methods</i></a>. (<i>A</i>) Cell density was accessed at different times after treatment and calculated as the number of live cells per cm² dish surface area. (<i>B</i>) Cell viability was determined by flow cytometry after 24 h and expressed as the percentage of PI and Annexin-V double-negative cells. (<i>C</i>) Immunocytochemistry detection of BrdU-positive nuclei (brown). (<i>D</i>) Representative histogram of BrdU positive cells assessed by flow cytometry (left), and respective quantification data (right). Data represent mean ± SEM of three independent experiments. §<i>p</i><0.05 and *<i>p</i><0.01 from respective control.