Fig. 3.

ESCRT-III is required for proper nuclear egress of HSV-1. a Confocal microscope images of HeLa and HeLa/CHMP4B KO cells treated with (left) control siRNA (siCt) or (Right) siRNAs to CHMP4A and CHMP4C (siCHMP4AC), respectively, for 48 h, and subsequently infected with HSV-1 for 22 h. Bars, 20 μm. Images are representative of three independent experiments. b Percent of cells (80–200 cells in each experiment) with aberrant punctate structures along with the nuclear rim in the experiment in (a). Data are shown as the mean ± SEM of three independent experiments. Electron microscope images of (c) HeLa and d HeLa/CHMP4B KO cells treated with siCt or siCHMP4AC, respectively, for 48 h and infected with HSV-1 for 22 h. Arrowheads indicate virions defective in the scission steps. C cytoplasm, N nucleus, NM nuclear membrane. Bar, 500 nm. Images are representative of three independent experiments. Percent of (e) perinuclear enveloped virions and f capsids in the cytoplasm of 14 cells in the experiments in (c, d). Data are shown as the mean ± SEM and are representative of three independent experiments. g–i HeLa and HeLa/CHMP4B KO cells treated with siRNA(s) as described in a were infected with HSV-1 at an MOI of (g) 10 or h 0.05, or with i influenza virus at an MOI of 0.01, and progeny virus titers were assayed at the indicated hours post infection (h.p.i). Data are shown as the mean ± SEM of three independent experiments. The indicated P-values were obtained using the unpaired Student’s t-test (b, e–i). n.s., not significant