FIG 8.

Gene silencing of GM3 synthase does not affect JCPyV infection in SVGA cells. SVGA cells were transfected with siRNAs targeted to the ST3GAL5 or control (scrambled) siRNA. At 72 h posttransfection, cells were either harvested for analysis of GM3 synthase mRNA expression (A) or infected (B). (A) RNA was extracted from 2 wells and reverse transcribed by reverse transcription-PCR (RT-PCR), and real-time PCR was performed with 1 μg cDNA using primers specific for GM3 synthase or GAPDH (housekeeping gene). Results are represented as the relative gene expression for triplicate real-time PCR samples relative to the GAPDH siRNA control. The percent reduction in gene expression relative to the scrambled control is expressed for each siRNA. (B) SVGA cells transfected with ST3GAL5 siRNAs or controls were infected with JCPyV or SV40. Infected cells were scored by indirect immunofluorescence. Data represent the average number of infected cells per visual field for four 10× fields of view for triplicate samples. Error bars indicate SD.