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Peptide from RNF213 truncation degrades ZIKV proteins and inhibits viral replication (A) The sequence of peptide from RNF213 truncation (PR-23) and control peptide. (B) A549 cells were infected with ZIKV (MOI = 0.01) and then treated with PR-23 (10 μM). Viral replication of cell supernatant was measured by plaque assay at 48 hpi. <t>(C–J)</t> <t>HEK293T</t> cells were treated with the indicated amount of PR-23 for 24 h, and then transfected with plasmids encoding HA-tagged ZIKV proteins. After 36 h, cell lysates were harvested and detected by western blot assay. GAPDH served as an internal control. (K) <t>Vero</t> cells were treated with the indicated dose of PR-23 for 6 h and transfected with the plasmid encoding GFP-tagged ZIKV E protein. After 36 h, the cells were fixed and subjected to immunofluorescence assay. The nucleus was stained with DAPI (blue). Images were obtained using a laser confocal microscope. Scale bars, 100 μm. Data are expressed as the mean ± SD from three independent experiments ( n = 3; ns, p > 0.05; ∗, p < 0.05; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001).
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Peptide from RNF213 truncation degrades ZIKV proteins and inhibits viral replication (A) The sequence of peptide from RNF213 truncation (PR-23) and control peptide. (B) A549 cells were infected with ZIKV (MOI = 0.01) and then treated with PR-23 (10 μM). Viral replication of cell supernatant was measured by plaque assay at 48 hpi. <t>(C–J)</t> <t>HEK293T</t> cells were treated with the indicated amount of PR-23 for 24 h, and then transfected with plasmids encoding HA-tagged ZIKV proteins. After 36 h, cell lysates were harvested and detected by western blot assay. GAPDH served as an internal control. (K) <t>Vero</t> cells were treated with the indicated dose of PR-23 for 6 h and transfected with the plasmid encoding GFP-tagged ZIKV E protein. After 36 h, the cells were fixed and subjected to immunofluorescence assay. The nucleus was stained with DAPI (blue). Images were obtained using a laser confocal microscope. Scale bars, 100 μm. Data are expressed as the mean ± SD from three independent experiments ( n = 3; ns, p > 0.05; ∗, p < 0.05; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001).
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Peptide from RNF213 truncation degrades ZIKV proteins and inhibits viral replication (A) The sequence of peptide from RNF213 truncation (PR-23) and control peptide. (B) A549 cells were infected with ZIKV (MOI = 0.01) and then treated with PR-23 (10 μM). Viral replication of cell supernatant was measured by plaque assay at 48 hpi. <t>(C–J)</t> <t>HEK293T</t> cells were treated with the indicated amount of PR-23 for 24 h, and then transfected with plasmids encoding HA-tagged ZIKV proteins. After 36 h, cell lysates were harvested and detected by western blot assay. GAPDH served as an internal control. (K) <t>Vero</t> cells were treated with the indicated dose of PR-23 for 6 h and transfected with the plasmid encoding GFP-tagged ZIKV E protein. After 36 h, the cells were fixed and subjected to immunofluorescence assay. The nucleus was stained with DAPI (blue). Images were obtained using a laser confocal microscope. Scale bars, 100 μm. Data are expressed as the mean ± SD from three independent experiments ( n = 3; ns, p > 0.05; ∗, p < 0.05; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001).
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Peptide from RNF213 truncation degrades ZIKV proteins and inhibits viral replication (A) The sequence of peptide from RNF213 truncation (PR-23) and control peptide. (B) A549 cells were infected with ZIKV (MOI = 0.01) and then treated with PR-23 (10 μM). Viral replication of cell supernatant was measured by plaque assay at 48 hpi. <t>(C–J)</t> <t>HEK293T</t> cells were treated with the indicated amount of PR-23 for 24 h, and then transfected with plasmids encoding HA-tagged ZIKV proteins. After 36 h, cell lysates were harvested and detected by western blot assay. GAPDH served as an internal control. (K) <t>Vero</t> cells were treated with the indicated dose of PR-23 for 6 h and transfected with the plasmid encoding GFP-tagged ZIKV E protein. After 36 h, the cells were fixed and subjected to immunofluorescence assay. The nucleus was stained with DAPI (blue). Images were obtained using a laser confocal microscope. Scale bars, 100 μm. Data are expressed as the mean ± SD from three independent experiments ( n = 3; ns, p > 0.05; ∗, p < 0.05; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001).
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ATCC african green monkey kidney cells vero
Peptide from RNF213 truncation degrades ZIKV proteins and inhibits viral replication (A) The sequence of peptide from RNF213 truncation (PR-23) and control peptide. (B) A549 cells were infected with ZIKV (MOI = 0.01) and then treated with PR-23 (10 μM). Viral replication of cell supernatant was measured by plaque assay at 48 hpi. <t>(C–J)</t> <t>HEK293T</t> cells were treated with the indicated amount of PR-23 for 24 h, and then transfected with plasmids encoding HA-tagged ZIKV proteins. After 36 h, cell lysates were harvested and detected by western blot assay. GAPDH served as an internal control. (K) <t>Vero</t> cells were treated with the indicated dose of PR-23 for 6 h and transfected with the plasmid encoding GFP-tagged ZIKV E protein. After 36 h, the cells were fixed and subjected to immunofluorescence assay. The nucleus was stained with DAPI (blue). Images were obtained using a laser confocal microscope. Scale bars, 100 μm. Data are expressed as the mean ± SD from three independent experiments ( n = 3; ns, p > 0.05; ∗, p < 0.05; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001).
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Peptide from RNF213 truncation degrades ZIKV proteins and inhibits viral replication (A) The sequence of peptide from RNF213 truncation (PR-23) and control peptide. (B) A549 cells were infected with ZIKV (MOI = 0.01) and then treated with PR-23 (10 μM). Viral replication of cell supernatant was measured by plaque assay at 48 hpi. (C–J) HEK293T cells were treated with the indicated amount of PR-23 for 24 h, and then transfected with plasmids encoding HA-tagged ZIKV proteins. After 36 h, cell lysates were harvested and detected by western blot assay. GAPDH served as an internal control. (K) Vero cells were treated with the indicated dose of PR-23 for 6 h and transfected with the plasmid encoding GFP-tagged ZIKV E protein. After 36 h, the cells were fixed and subjected to immunofluorescence assay. The nucleus was stained with DAPI (blue). Images were obtained using a laser confocal microscope. Scale bars, 100 μm. Data are expressed as the mean ± SD from three independent experiments ( n = 3; ns, p > 0.05; ∗, p < 0.05; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001).

Journal: iScience

Article Title: RNF213 isoform 2 restricts Zika virus through antiviral signaling and viral protein degradation

doi: 10.1016/j.isci.2026.116284

Figure Lengend Snippet: Peptide from RNF213 truncation degrades ZIKV proteins and inhibits viral replication (A) The sequence of peptide from RNF213 truncation (PR-23) and control peptide. (B) A549 cells were infected with ZIKV (MOI = 0.01) and then treated with PR-23 (10 μM). Viral replication of cell supernatant was measured by plaque assay at 48 hpi. (C–J) HEK293T cells were treated with the indicated amount of PR-23 for 24 h, and then transfected with plasmids encoding HA-tagged ZIKV proteins. After 36 h, cell lysates were harvested and detected by western blot assay. GAPDH served as an internal control. (K) Vero cells were treated with the indicated dose of PR-23 for 6 h and transfected with the plasmid encoding GFP-tagged ZIKV E protein. After 36 h, the cells were fixed and subjected to immunofluorescence assay. The nucleus was stained with DAPI (blue). Images were obtained using a laser confocal microscope. Scale bars, 100 μm. Data are expressed as the mean ± SD from three independent experiments ( n = 3; ns, p > 0.05; ∗, p < 0.05; ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001).

Article Snippet: Human embryonic kidney (HEK293T), human lung adenocarcinoma (A549), and African green monkey kidney (Vero) cells were purchased from the American Type Culture Collection (ATCC), IFNAR-knockout A549 cells were kindly provided by Prof. Genhong Cheng (Guangzhou National Laboratory).

Techniques: Sequencing, Control, Infection, Plaque Assay, Transfection, Western Blot, Plasmid Preparation, Immunofluorescence, Staining, Microscopy