Depletion of PML or Sp100 does not interfere with latency establishment in HUVEC cells.

<p>(<b>A</b>) Western blot analysis of HUVEC cells transduced with shRNA-expressing lentiviruses directed against PML, Daxx, Sp100 or GFP. shDaxx transduced cells did not show a reduction of Daxx protein levels and were thus not included in further analyses. Although residual levels of PML and Sp100 are still visible in bulk protein extracts, immunofluorescence analysis suggests that a considerable number of cells is devoid of PML or Sp100 positive nuclear bodies (<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004274#ppat.1004274.s009" target="_blank">Figure S9</a>). (<b>B</b>) FACS analysis of stably shRNA-expressing HUVEC cultures analysed 48 h ater infection with KSHV. The rightmost columns show cells which were treated with sodium butyrate (5 mM) immediately after infection. Mock infected cells were used to correct for background staining levels. Error bars represent SEM of three biological replicates. (<b>C</b>) Transcript levels of ORF50, ORF59, ORF64 and ORF73 in EA.hy cells at 48 hours post infection. Expression was calculated by normalization to GAPDH and is shown relative to shGFP controls (set to 1). Error bars represent SEM of at least three data sets.