FIG 12.

E-Syt1 and E-Syt3 impair HSV-1 entry. HeLa cells seeded in 6-well plates were transfected with siRNAs targeting E-Syt1, E-Syt3, or both for 48 h. Control nontargeting siRNAs (NC1) were also used. The cells were subsequently mock infected or infected at an MOI of 30 for 1 h at 4°C (viral adsorption step) with wild-type HSV-1 (luciferase negative) or HSV-1 Lox-Luc (luciferase positive). The cells were then incubated at 37°C for another hour to enable viral entry and subsequently lysed at room temperature for 30 min in the presence of luciferin and energy. As an additional control, one sample was left at 4°C throughout the experiment to prevent viral entry. All the samples were next transferred to 96-well plates and analyzed with a luminometer. The values represent the mean relative light units (RLU) from three independent experiments, and the error bars indicate the standard errors of the mean. The asterisks indicate the results of bilateral Student's t tests upon comparison to the HSV-1 Lox-Luc positive control. The following P values were obtained: mock treated, 8 × 10⁻⁹; HSV-1 17⁺, 1 × 10⁻⁹; 4°C, 0.0004; NC1, 0.86; siE-Syt1, 0.001; siE-Syt3, 0.028; siE-Syt1 and -3, 0.024 (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant).