Fig. 8: Impaired phospholipid remodeling reduces viral particle production but does not affect virus entry or RNA replication and protein expression.

a shRNA-transduced Huh7 cells were infected at 4 dpt with high MOIs (ZIKV MOI 2, WNV MOI 5, DENV MOI 2, TBEV MOI 10, and YFV-17D MOI 5). Virus entry was determined by measuring viral genome equivalents (GE) 4 h post infection using RT-qPCR (mean ± SEM, technical duplicates from 2 to 3 independent experiments, non-significant in unpaired two-tailed Welch’s t-test). b, c Transduced Huh7 cells were infected with low MOIs 4 dpt (ZIKV MOI 0.1, WNV MOI 0.001, TBEV MOI 0.05, DENV MOI 0.05, and YFV-17D MOI 0.005) and analyses were performed at 2 dpi. b Intracellular viral RNA (vRNA) was quantified using RT-qPCR (mean ± SEM, n = 4 (DENV), or n = 5 (ZIKV, TBEV, WNV, YFV) independent experiments, non-significant in unpaired two-tailed Welch’s t-test) and the viral envelope/E protein level was determined by immunoblot analysis. Box-and-whisker plot indicates E protein expression as log₂ fold change over shNT (center line: median, box limits: upper and lower quartiles, whiskers: 1.5 × interquartile range, points: outliers, n = 3 (TBEV, YFV-17D) or n = 4 (ZIKV, WNV, DENV) independent experiments, non-significant in unpaired two-tailed one sample Student’s t-test). c Extracellular vRNA was quantified by RT-qPCR (mean ± SEM, n = 4 (ZIKV, YFV-17D) or n = 5 (TBEV, WNV, DENV) independent experiments, non-significant in unpaired two-tailed Welch’s t-test) and E protein levels in the supernatant by immunoblot analysis. Box-and-whisker plot indicates E protein levels as log₂ fold change over shNT (center line: median, box limits: upper and lower quartiles, whiskers: 1.5 × interquartile range, points: outliers, n = 2 (YFV-17D) or n = 5 (ZIKV, WNV, TBEV) independent experiments, *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 unpaired two-tailed one sample Student’s t-test). dpt days post transduction, dpi days post infection. Source data are provided as a Source Data file.