FIG 7.

Lipoquads inhibit HCV cell-to-cell transmission. Huh7/Scr cells were infected with Jc1 HCVcc at an MOI of ∼10 TCID₅₀/cell and 20 h later seeded on 24-well plates. Simultaneously, Huh7.5/EGFP-NLS-IPS cells were seeded on the same 24-well plates (coculture, at a ratio 1:1 with Huh7/Scr cells), and lipoquads (10 μM) or dasatinib (10 μM) was added to the wells. The addition of the compound solvent (lipoquad solvent or DMSO) served as a negative control. Four hours later, medium was removed and cells were overlaid with 1% agarose with fresh compounds. Twenty-four hours later, the HCV infection was analyzed by immunofluorescent NS5A staining. (A) Results are the means of quantification of Huh7.5/EGFP-NLS-IPS cells in 3 independent wells by taking 3 independent pictures of different fields of each well that contained at least 200 cells in total. Cell-to-cell inhibition is expressed as the percentage of acceptor cells with the EGFP-NLS plus NS5A signal with respect to the total number of EGFP plus NS5A cells in the DMSO-treated (for dasatinib) or lipoquad buffer-treated (for lipoquads) wells. (B) Representative images of coculture cells stained with anti-NS5A antibodies. Huh7.5/EGFP-NLS-IPS cells with EGFP signal redestributed to the nucleus and cytoplasmatic NS5A signal are indicated with arrows. Magnification, ×63.