FIG. 8.

CD36⁺/Epo⁺ EPCs treated with Jak2-specific shRNA show reduced susceptibility to B19V infection. CD36⁺/Epo⁺ EPCs at day 6 of culture were transduced with Lenti-GFP-Jak2-shRNA and Lenti-GFP-Scramble-shRNA. They were infected with B19V 48 h later (day 8) and analyzed at 48 h p.i. (A) Expression levels of Jak2 and B19V NS1 in GFP-expressing cells were selectively gated to quantify the expression levels of Jak2 and B19V NS1 by flow cytometry (left) and quantification thereof (right). Histograms indicate the average percentage of B19NS1-expressing cells in each group, and quantification is based on the MFI of Jak2 expression. Bkg, secondary antibody-only staining background. (B and C) Cells were harvested for mRNA isolation using the Turbo mRNA kit, as described in Materials and Methods. RT-qPCR was carried out to assess the B19V VP2-encoding mRNA/β-actin mRNA (B) and the B19V D1/A1-2-spliced mRNA/β-actin mRNA (C). Histograms illustrated in panels B and C show the averages and standard deviations and were calculated from at least three independent experiments.