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. 2015 Jan 15;475:66–73. doi: 10.1016/j.virol.2014.11.007

Fig. 1.

Fig. 1

A reduction in RAB1A expression inhibits VACV replication. (A) Western blot image of protein lysates from HeLa cells mock transfected or transfected with non-targeting siRNA (RSCF) or a siRNA SMARTpool targeting RAB1A, probed with antibody against RAB1A (green) or actin (red), and visualised using the Odyssey® Infrared Imaging System (Li-Cor Biosciences). (B) HeLa cells (seeded on 96 well plates) were mock transfected or transfected in triplicate with non-targeting siRNA (RSCF), or siRNA targeting PRK-AB1, a RAB1A siRNA SMARTpool (SP), or four individual deconvoluted RAB1A siRNAs (DC1–DC4). After 48 h, cells were infected with VACV-A5L-EGFP at MOI=0.1, and after a further 48 h fluorescence levels were measured. Values shown are the mean corrected for background, error bars represent standard error of the mean, and P values <0.05 are indicated (t-test). (C, D) Multistep growth curves. HeLa cells were mock transfected or transfected with non-targeting siRNA (VP16) or a siRNA SMARTpool targeting RAB1A for 48 h, then infected with a MOI of 0.1 with either (C) VACV-A5L-GFP and virus levels determined by fluorescence, or (D) VACV and the amount of infectious virus levels in both the supernatant and cells combined determined by plaque assay on BS-C-1 cells. (C) Means of three technical replicates, error bars indicating the standard deviation. P values <0.05 are indicated (t-test). The data are representative of three biological replicates. (D) Average and standard error of the mean (SEM) of five biological replicates. P values <0.05 are indicated (mixed model analysis).