Fig. 2.

Pretreatment of Wolbachia-infected Aag2 cells with low concentrations of 2-hydroxypropyl-β-cyclodextrin (2HPCD) reverses inhibition of DENV replication. a Uninfected cells (–ve) and wMelPop-infected cells were treated for 48 h with various concentrations of 2HPCD prior to addition of DENV virus at a multiplicity of infection of 0.1. Cells were harvested 5 days post infection and DENV levels measured by qPCR. Significant differences of <0.05 are indicated as determined by analysis of variance (ANOVA) corrected by Tukey’s HSD test, n = 5. b Cholesterol turnover imaged using TopFluor cholesterol. Uninfected and wMelPop-infected cells were pretreated with either PBS or 0.1 mM 2HPCD for 24 h. Cells were then labelled with TopFluor cholesterol for 30 min and subsequently incubated with either PBS or 0.1 mM 2HPCD for 24 h before imaging by fluorescence confocal microscopy; scale bar indicates 10 µm. Lipid droplets were quantified as TopFluor-positive spots/cell; error bars denote s.d. Statistical significance was assessed by ANOVA corrected by Tukey’s HSD test, n = 5. c Imaging of Nile Red stained or TopFluor-treated wMelPop cells and measurement of lipid droplet size