FIG 1.

Effect of increasing artesunate concentrations on BKV replication in RPTECs. (A) Extracellular BKV DNA load. At 72 hpi, supernatants were harvested from BKV-infected RPTECs treated with artesunate at the indicated concentrations from 2 hpi, and BKV DNA loads were measured by qPCR. Mean values of the number of GEq/ml ± SDs of four experiments (each experiment was performed in two wells) are presented. *, P <0.05, determined by the t test. (B) Expression of BKV proteins. At 72 hpi, BKV-infected RPTECs treated with the indicated concentrations of artesunate from 2 hpi were fixed and stained with polyclonal rabbit anti-agnoprotein serum for visualization of the BKV late agnoprotein (green) and with the SV40 LT-ag monoclonal antibody PAb416 for visualization of the BKV early protein LT-ag (red). Cell nuclei (blue) were stained with Draq5. The pictures were taken with a fluorescence microscope (×10 objective). (C) Infectious progeny release. The supernatants from untreated and artesunate-treated BKV-infected RPTECs harvested at 72 hpi were also seeded onto new RPTECs. At 72 hpi, these cells were fixed and indirect immunofluorescence staining was performed, as described for panel B. The pictures were taken with a fluorescence microscope (×10 objective).