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. 2010 Sep 22;84(23):12226–12235. doi: 10.1128/JVI.00994-10

FIG. 3.

FIG. 3.

PV induces Ca2+ release from the ER through IP3R and RyR channels in IMR5 cells. (A) Smaller increase in the cytosolic Ca2+ concentration ([Ca2+]c) in PV-infected IMR5 cells treated with the IP3R channel inhibitor 2-APB or the RyR inhibitor dantrolene. Cells were mock infected or infected with PV in medium without Ca2+ (light gray) or in medium with Ca2+ in the presence of 10 μM 2-APB (dark gray) or 20 μM dantrolene (gray) or in the absence of inhibitor (black). At the indicated times p.i., cells were analyzed by flow cytometry after FLUO3-AM staining. The increase (n-fold) in cytosolic Ca2+ was calculated as the ratio of the percentage of fluorescent PV-infected IMR5 cells to the percentage of fluorescent mock-infected cells. The data shown are the means from three independent experiments. Error bars indicate the standard errors of the means. *, P < 0.05 by Student's t test comparing untreated IMR5 cells to treated IMR5 cells. (B) The treatment of IMR5 cells with 2-APB, XeC, or dantrolene does not affect PV growth. IMR5 cells were infected with PV for 8 h in the presence or absence of 2-APB (10 μM), XeC (10 μM), or dantrolene (20 μM). Total virus yield (extracellular and intracellular) was determined by TCID50 assay after three cycles of freezing and thawing to release intracellular viruses. Each point represents the mean virus titers for three independent experiments. Error bars indicate the standard errors of the means. (C) Smaller increase in cytosolic Ca2+ concentration ([Ca2+]c) in PV-infected IMR5 cells treated with the IP3R channel inhibitor XeC. Mock-infected and PV-infected (8 h p.i.) IMR5 cells treated with 10 μM XeC (gray) or left untreated (black) were analyzed at the indicated times p.i. by flow cytometry after FLUO3-AM staining. The increase (n-fold) in cytosolic Ca2+ was calculated as the ratio of the percentage of fluorescent PV-infected IMR5 cells to the percentage of fluorescent mock-infected cells. The data shown are the means from three independent experiments. Error bars indicate the standard errors of the means. *, P < 0.05 by Student's t test comparing untreated IMR5 cells to treated IMR5 cells.