Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Heparan sulfate as an attachment factor for ASFV is restricted by host HB-EGF

doi: 10.1007/s00018-026-06188-z

Figure Lengend Snippet: Host HB-EGF inhibits ASFV infection by blocking viral attachment and internalization in vitro. A PAMs were infected with purified ASFV-WT (MOI = 2) at 37 °C for 0, 6, 12, and 24 h, respectively. The HB-EGF mRNA expression levels were detected by qPCR. MA-104 cells were transfected with HB-EGF for 24 h, followed by infection with ASFV-WT (MOI = 0.1) at 37 °C for 24 h. The ASFV infection was detected by western blot ( B ) and qPCR ( C ). HB-EGF stable expression MA-104 cells were infected with ASFV-WT (MOI = 0.1) at 37 °C for 24 h. The ASFV infection was detected by western blot ( D ) and qPCR ( E ). HB-EGF-KD MA-104 cells were infected with ASFV-WT (MOI = 0.1) at 37 °C for 24 h. The ASFV infection was detected by western blot ( F ) and qPCR ( G ). The attachment ( H ) and internalization ( I ) of ASFV in Wild-type, HB-EGF stable expression, and HB-EGF-KD MA-104 cells were detected by qPCR. The attachment ( J ) and internalization ( K ) of DiD-ASFV were observed by ZEISS LSM980. The cell membrane stained with green markers was used as a reference, where the cell membrane was colocalized (red) with the binding virus. The attachment ( L ) and internalization ( M ) were counted. More than 100 cells were counted per sample, and the number of cells that bound DiD-ASFV was calculated

Article Snippet: Surfen dihydrochloride, mitoxantrone, heparin, heparan sulfate, DiD perchlorate, and recombinant HB-EGF eukaryotic expression protein were purchased from MedChemExpress (USA).

Techniques: Infection, Blocking Assay, In Vitro, Purification, Expressing, Transfection, Western Blot, Membrane, Staining, Binding Assay, Virus

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Heparan sulfate as an attachment factor for ASFV is restricted by host HB-EGF

doi: 10.1007/s00018-026-06188-z

Figure Lengend Snippet: HB-EGF inhibits ASFV infection by the Heparin Binding domain. Schematic diagram of HB-EGF domain structure ( A ) and the truncation mutants ( B ) used to determine the antiviral domain. MA-104 cells were transfected with HB-EGF-Full, HB-EGF-ΔED, and HB-EGF-ΔHBD, followed by infection with ASFV-WT (MOI = 0.1) at 37 °C for 24 h. The ASFV infection was detected by western blot ( C ) and qPCR ( D ). Schematic diagram of HBD-derived peptide sequence ( E ). The affinity of the peptide for heparin ( F ) and heparan sulfate ( G ) was detected by Bio-Layer Interferometry. PAMs were pretreated with different concentrations of peptide, followed by infection with ASFV-WT (MOI = 0.1) at 37 °C for 24 h ( H ). ASFV p72 protein and mRNA expression were assessed by western blot ( I ) and qPCR ( J ). The progeny virus production was quantified by the HDA50 assay ( K ). Recombinant HB-EGF (200 ng/ml) was used to pretreat PAMs at 37 °C for 6 h in the presence or absence of Heparin (200 ng/ml), followed by infection with ASFV-WT/GFP (MOI = 0.1) at 37 °C for the indicated time points ( L ). ASFV infection was quantified by western blot ( M ), fluorescence microscopy ( N , O ), and HAD50 assay ( P )

Article Snippet: Surfen dihydrochloride, mitoxantrone, heparin, heparan sulfate, DiD perchlorate, and recombinant HB-EGF eukaryotic expression protein were purchased from MedChemExpress (USA).

Techniques: Infection, Binding Assay, Transfection, Western Blot, Derivative Assay, Sequencing, Expressing, Virus, Recombinant, Fluorescence, Microscopy