FIG 5.

SFK inhibitors do not reduce CHIKV RNA synthesis or replication complex formation. (A and B) NHDFs were infected with CHIKV_181/25 (A) or VEEV (B) (MOI = 3 PFU/cell). At 2 hpi, cells were washed twice with PBS, and cells were treated with 10 μM dasatinib. At 12 hpi, cells were washed extensively in PBS, and cells were lysed with TRIzol reagent for total RNA isolation. Relative levels of vRNA were analyzed by qRT-PCR using gene-specific primers and probes directed against CHIKV E1 and VEEV E2. Statistical analyses were performed on log-transformed data, data were analyzed by Dunn’s multiple-comparison test, and multiplicity-adjusted P values are reported (n = 3) (*, P < 0.05). (C) RNAs from uninfected and CHIKV_181/25-infected cells (MOI = 3 PFU/cell) treated with and without dasatinib were analyzed by Northern blotting for genomic and subgenomic RNA levels at 12 hpi. (D) NHDFs were infected with CHIKV_181/25 (MOI = 25 PFU/cell) and treated with dasatinib at 2 hpi or left untreated (ND). At 8 hpi, cells were fixed and stained for dsRNA (J2) (green), actin (phalloidin) (red), and DNA (DAPI) (blue).