Fig. 8

Addition of mannose can reverse the negative impact of 2-DG on the LCMV life cycle. MRC-5 cells were infected with LCMV at an MOI of 3. After adsorption, the inoculum was removed and replaced with fresh media according to experimental design, followed by 24-h incubation. A, B Whole-cell extracts were harvested and used for the immunoblot analysis of viral protein expression levels using anti-GP2 (A), anti-NP, and anti-Z antibodies (B). The signals obtained with anti-α-tubulin or anti-β-actin antibodies were used as loading controls. The ratios of GP-C/tubulin, GP2/tubulin, NP/actin, and Z/actin were determined based on the densitometric analysis of protein abundance. Final values are expressed relative to control, which was set to 1. C LCMV titers in the supernatants were analyzed by FFA. Data from six independent experiments are shown as a scatter of individual values. Mean and SD are shown as column and error bars, respectively. D Total cellular RNA was isolated and LCMV RNA was detected by RT-qPCR. Copy numbers of viral NP were calculated using a standard curve. Data from three independent experiments are shown as a scatter of individual values. Mean and SD are shown as column and error bars, respectively. Statistical differences between groups were analyzed using Welch’s ANOVA with Tamhane’s T2 post hoc test (C) or one-way ANOVA with Tukey’s post hoc test (D). Different letters above the columns indicate significant differences (P ≤ 0.05), while identical letters indicate no significant difference. E Whole-cell extracts were subjected to either mock or PNGase F treatment and used for the immunoblot analysis of viral glycoprotein levels using anti-GP2 antibody. The signal obtained with anti-β-actin antibody was used as a loading control. NP, nucleoprotein; Z, Z protein; glyco/unglyco GP-C, glycosylated/unglycosylated glycoprotein precursor complex; GP2, glycoprotein 2; glyco/unglyco GP2, glycosylated/unglycosylated glycoprotein 2; FFU/ml, focus-forming units per milliliter