FIG 4 .

Independent of entry, Wolbachia blocks viral replication. (A and B) Wolbachia wStri-infected cells (W+) and Wolbachia-free cells (W−) cells were transfected with 1 µg of ZIKV UTR probe with a AlexaFluor 547 conjugate (red) to assess transfection efficiency differences. (A) Cells were fixed and counterstained with DAPI (DNA; blue). Control cells received probe only without transfection reagent or reagent only without probe. (B) Fluorescence was detected by using a Tecan Spark plate reader at 547 nm. (C) ZIKV RNA was isolated by Trizol extraction, and 0.5 or 1 µg of viral RNA was transfected into W− or W+ cells. Four days posttransfection, supernatant was collected and virus production was quantified by plaque assay. The data shown are means and standard deviations of results from three independent experiments. Statistical significance was determined by using Student’s t test. *, P < 0.05.