Fig. 4: EDC4 levels impact viral RNA production and EBOV growth kinetics.

a EDC4 protein levels after siRNA treatment was measured by immunoblotting with EDC4-specific antibodies. Lower panel shows quantitation of a representative replicate of three independent experiments measured by area under the curve and normalized to β-actin expression levels. b EDC4 was depleted using the indicated siRNA and samples were harvested at 2, 6, 18, 24, 48, and 72 hpi. EBOV was titered by FFU assays performed on VeroE6 cells. Three technical replicates of one biological replicate are shown. Statistical significance was calculated by one-way ANOVA. Data are presented as mean ± SD. P-values = * 0.0271; ** 0.0051. c Viral RNA synthesis was measured by RNAFISH staining using multiple oligonucleotides complementary to EBOV NP and VP35 mRNA. Scale bar = 100 μm. d Quantification of RNAFISH signal from 10 images for which panel c is representative. Two technical replicates of one representative biological replicate are shown. e Measurement of viral RNA species by qPCR. NP and GP mRNA levels were measured after cDNA synthesis using poly-dT primers. Genomic (negative sense) and anti-genomic (positive sense) RNA were measured using a two-step reaction with specific primers (see methods). One representative biological replicate is shown per qPCR, each containing two or three technical replicates as indicated. Source data are provided as a Source Data File.