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. 2021 Aug 2;40(18):e105658. doi: 10.15252/embj.2020105658

Figure 6. Functional interaction between VP30 and multiple host proteins that contain PPxPxY motifs.

Figure 6

  1. The effect of over‐expression of the indicated host proteins on EBOV RNA synthesis was assessed using an EBOV MG assay. HEK293T cells were transfected with plasmids expressing components of the MG assay and either 12.5 or 125 ng of expression plasmids for the indicated host factors. Empty expression plasmid (vector) served as a control. Luciferase activity was measured 48 h post‐transfection and fold MG activity was calculated relative to no VP30 control.
  2. MG activity upon transient expression of GFP or GFP‐fused PPxPxY‐containing peptides. Plasmids expressing GFP‐fused peptides were transfected at 12.5 and 125 ng amounts. Statistical significance was calculated relative to the GFP control for each concentration.
  3. MG activity upon knockdown of host genes. HEK293T cells were transfected with scrambled siRNA or siRNA targeting RBBP6, hnRNP L, hnRNPUL1, or PEG10. Twenty‐four hours post‐transfection, cells were transfected with MG assay plasmids including VP30 at 12.5 and 25 ng doses.
  4. MG activity was assessed as in panel C upon single or double knockdown of RBBP6 and hnRNP L.

Data information: For each panel (A–D), the data represent the mean ± SD from one representative experiment in which each transfection condition was performed in triplicate (n = 3). The experiment was reproduced in two additional, independent experiments (see Appendix Figs S3, S4, S8 and S9). Statistical significance was calculated using ANOVA with Tukey’s multiple comparisons test. ****P < 0.00005; ***P < 0.0005, **P < 0.005, *P < 0.05.