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From:
Published online 2010 April 15. doi: 10.1371/journal.ppat.1000847

Figure 6

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The role of the IFN β pathway in platelet production and JUNV infection.

(A) CD34+ cells were treated with poly(I:C) (100 µg/ml) at the indicated days, stimulated with TPO, and Plt counts were determined at day 15 of the culture. The values represent the mean ± SEM of four independent experiments, * p<0.05 vs. vehicle (control). (B) IFN β mRNA levels in UV-irradiated JUNV- or JUNV-infected CD34+ cells were determined by RT-PCR at the indicated days of culture. The figure shows a representative experiment of three similar replicates. (C) CD34+ cells were treated with IFN β and stimulated with TPO. Plt counts were determined at day 15 of the culture. The values represent the mean ± SEM of three independent experiments,* indicates p<0.05 vs. no IFN β, # indicates p<0.05 vs. IFN β (10 U/ml). (D) Total cell number was determined at the indicated days of culture by counting cells with a hemocytometer. Similar results were obtained in MTT assays. The values represent the mean ± SEM of three independent experiments. (E) Anti-IFN β (1,000 neutralizing units) or an equal volume of rabbit Igs was added before CD34+ cell infection and Plt counts were determined at day 15. Values represent the mean ± SEM of three independent experiments, * indicates p<0.05 vs. UV-irradiated JUNV. (F) Type I IFN receptor subunit (IFNAR1 and 2) mRNAs were evaluated by RT-PCR in megakaryocyte precursors purified by immunomagnetic positive selection (99±1% of purity). The figure shows a representative experiment of three similar replicates. (G) CD34+ cells were infected with JUNV and 1000 neutralizing units of anti-IFN β or an equal volume of rabbit Igs were added before TPO stimulation. Viral antigens were detected by flow cytometry. The figure shows a representative experiment of two similar replicates. (H) The kinetics of TfR1 expression in the presence or absence (vehicle) of IFN β (10 U/ml) in CD34+ cells stimulated with TPO were determined by flow cytometry.

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