Journal: Journal of Virology

Article Title: Role of g5Rp in African swine fever virus replication: disruption of host translation and autophagy

doi: 10.1128/jvi.01252-25

Figure Lengend Snippet: g5Rp interacts with eIF5A and RPS15, modulating host translation to enhance ASFV replication. ( A, B ) Co-IP of ASFV-infected lysates showing g5Rp interaction with eIF5A ( A ) and RPS15 ( B ). ASFV infection at MOI of 1 for 24 hpi. ( C, D ) PLA indicating g5Rp interaction with eIF5A ( C ) and RPS15 ( D ) in ASFV-infected cells. ( E, F ) Validation of g5Rp interaction with eIF5A ( E ) and RPS15 ( F ) by co-transfection in HEK293T cells, immunoprecipitation with anti-Flag, and immunoblotting with anti-HA. ( G, H ) Viral titers measured by TCID50 assay in cells with knockdown or overexpression of eIF5A ( G ) and RPS15 ( H ). ( I, J ) Western blotting analysis showing p30 expression in eIF5A ( I ) knockdown and RPS15 ( J ) overexpression conditions at 24 hpi. β-actin was the loading control. Data are represented as mean ± SD of three independent experiments. Statistical significance was determined using Student’s t -test: *, P < 0.05; **, P < 0.01; ns, not significant.

Article Snippet: Cell lysates were centrifuged (12,000 × g , 10 min), and supernatants were pre-cleared by incubation with protein A/G agarose (Sigma) at 4°C for 1 h. The pre-cleared supernatants were then incubated with anti-Flag (M2) agarose beads or with protein G plus agarose beads (MedChemExpress) pre-bound with 1 μg of control IgG antibody overnight at 4°C on a roller.

Techniques: Co-Immunoprecipitation Assay, Infection, Biomarker Discovery, Cotransfection, Immunoprecipitation, Western Blot, TCID50 Assay, Knockdown, Over Expression, Expressing, Control