Fig. 3: SOAT1 inhibitor treatment, but not DGAT inhibitors reduce ZIKV infection in Huh7 cells.

a Scheme of the experimental setup. Huh7 cells were pre-treated with DGAT inhibitors (5 µM each), SOAT1-specific inhibitors (ATR-101 = 6 µM or 9.72 µM, K604 = 5 µM, 10 µM, or 20 µM), or DMSO 24 h prior to infection with ZIKV (MOI 0.1). After 1 h, the inoculum was removed and replaced by fresh media supplemented with the respective inhibitors. At 48 hpi, supernatants were harvested and cells were lysed. b Cell lysates and supernatants were analyzed by immunoblotting using ZIKV C and orthoflavivirus E-protein specific antibodies (Lys = cell lysates, Sup = supernatant). GAPDH served as loading control. Shown is one representative immunoblot. Bands were quantified with ImageLab and intracellular signals were normalized to GAPDH. Box-and-whisker plot shows viral protein level as log₂ fold change over DMSO control (center line: median, box limits: upper and lower quartiles, whiskers: 1.5 x interquartile range, points: outliers, n = 3, except for K604 5 µM and 10 µM (n = 2), *p ≤ 0.05, **p ≤ 0.01, no asterisk = not significant, one-sample t-test). c Supernatants from cells in (b) were analyzed using TCID₅₀ titration (mean ± SEM, n = 3, no asterisk = not significant, Welch´s t-test). d After 4 d, cells were fixed, surviving cells were visualized by crystal violet staining, and CPE was quantified using Fiji. Box-and-whisker plot indicates CPE as log₂ fold change over DMSO control (center line: median, box limits: upper and lower quartiles, whiskers: 1.5 x interquartile range, points: outliers, n = 3, except for K604 5 µM and 10 µM (n = 2), *p ≤ 0.05, **p ≤ 0.01, no asterisk = not significant, one-sample t-test).