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. 2010 Sep 20;120(10):3530–3544. doi: 10.1172/JCI41805

Figure 2. B19V NS1 protein regulates the E2F family of transcription factors.

Figure 2

(A) Immunoblot analysis of E2F transcription factors. CD36+ EPCs were B19V- or mock-infected or transduced with NS1 or control lentivirus. Whole cell lysates were prepared at the indicated time points, resolved on 4%–12% SDS-PAGE, and subjected to immunoblot analysis with antibodies specific for individual E2Fs, B19V VP2, Flag (NS1), or GAPDH. (B) Transcription profiles of E2F target genes affected by NS1. CD36+ EPCs were transduced with NS1 or control lentivirus, followed by total RNA isolation at the indicated time points and then microarray analysis using Affymetrix HG-U133 Plus 2 arrays as described in Methods. Among 1,045 genes with greater than 1.5-fold changes, 93 were identified as E2F targets. Among them, 72 genes were involved in cell cycle regulation (44 upregulated and 28 downregulated) and 21 genes (8 upregulated and 13 downregulated) in cell differentiation. The heat map represents normalized intensity (NS1 divided by control) for each time point. Gene symbols are listed to the left of each panel.