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. 2014 Oct 13;9(10):e109969. doi: 10.1371/journal.pone.0109969

Figure 2. Absence of HCV infection by Ca-alg encapsulated cells.

Figure 2

(A) Encapsulated cell cultures were established by encapsulating HuH-7-RFP-NLS-IPS cells within Ca-alg beads (500 000 cells/mL). Following the cell encapsulation stage, 400 µL of beads were transferred to tissue culture 6-well plates with 1 mL of DMEM medium added on 3D moving plates. After 4 h of contact with the JFH1 virus (HCV+) or without (HCV), the medium was removed and replaced with 2 mL of fresh complete DMEM medium. Ca-alg beads devoid of HuH-7 cells were used as controls. (B) Foci of infected cells (in 2D or in beads), identified by translocation of the cleavage product RFP-NLS from cytoplasm to nucleus, were visualized at 48 h by fluorescence microscope. Images are representative of three independent experiments. Nuclei were stained by DAPI. (C) The amount of HCV RNA was quantified in the bead supernatants by RT-qPCR. Results are expressed as HCV RNA IU/mL and are reported as the mean ± S.D. of triplicate measurements. (D) Viral titers were determined in the bead supernatants by FFU assay. Results are expressed as FFU/mL and are reported as the mean ± S.D. of three independent experiments. ***P<0.0001, ns: no significant difference.