ORF45 reduces cell adhesion through RSK activation.
<p>A. HEK293 cells were infected with IRES-zsGreen empty vector, ORF45 WT or F66A-expressing lentiviruses (MOI = 2). Forty-eight hours later, the cells were dissociated and seeded on a fibronectin-coated 96-well plate. After 1 h incubation, the unattached cells were removed by low shear washin...
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2025
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| Resumo: | <p>A. HEK293 cells were infected with IRES-zsGreen empty vector, ORF45 WT or F66A-expressing lentiviruses (MOI = 2). Forty-eight hours later, the cells were dissociated and seeded on a fibronectin-coated 96-well plate. After 1 h incubation, the unattached cells were removed by low shear washing, the attached cells were observed and recorded under fluorescence microscopy, and a representative image was shown. B. HEK293 cells transfected with ORF45 and RSK2 WT- or mutant-expressing plasmids were dissociated and seeded on a fibronectin-coated 96-well plate, and the relative focal adhesion was calculated and shown as mean ± standard deviation (SD). *, p < 0.05; **, p < 0.01; t test. C. Cell detachment is induced during lytic replication. iSLK.219 cells were seeded on SLK cell monolayers at a 1:50 ratio and then induced with Dox and NaB for 48 h. The living cells were fixed and directly visualized with confocal fluorescence microscopy. D. iSLK-Bac16 cells and iSLK-STOP45 cells were induced with Dox and NaB, or iSLK.STOP45 cells were infected with ORF45 WT or F66A-expressing lentiviruses (MOI = 5) and then cells were induced with Dox and NaB at 12 h post-infection. The cells were visualized with inverted microscopy at 72 h post-induction. The quantification of relative cell adhesion was calculated and shown. **, p < 0.01; t test.</p> |
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