Filamin A phosphorylation is induced by ORF45-mediated RSK activation.
<p>A. Ectopic ORF45 WT and F66A, RN or RC mutated constructs were introduced into SLK cells by infection with recombinant lentiviruses. The cells were collected at 72 h post-infection, and whole cell lysates were analyzed by Western Blots as indicated. B. HEK293 cells were cotransfected with H...
সংরক্ষণ করুন:
| প্রধান লেখক: | |
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| অন্যান্য লেখক: | , , , , |
| প্রকাশিত: |
2025
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| বিষয়গুলি: | |
| ট্যাগগুলো: |
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| সংক্ষিপ্ত: | <p>A. Ectopic ORF45 WT and F66A, RN or RC mutated constructs were introduced into SLK cells by infection with recombinant lentiviruses. The cells were collected at 72 h post-infection, and whole cell lysates were analyzed by Western Blots as indicated. B. HEK293 cells were cotransfected with HA-RSK2 WT-, CA- or KD-expressing plasmids with empty vector or ORF45-expressing plasmids for 24 h, and then, the cells were serum-starved overnight, collected and subjected to western blot analysis as indicated. C. iSLK- cells and iSLK.219 cells were induced with 1 µg/ml Dox and 1 mM NaB. The cells were collected at different time points, and whole cell extracts were detected as indicated. D. Control cells, iSLK-Bac16, -STOP45, ORF45F66A, ORF45RN or ORF45RC cells were induced with Dox and NaB as described above, the cells were collected at 72 h after induction, and whole cell extracts were analyzed. E. HEK293 cells were infected with Bac16 or STOP45 virions (MOI = 10). At the different time points post-infection, the cells were collected and whole cell extracts were prepared and subjected to Western Blotting analysis as indicated. F. The iSLK.Bac16 or iSLK.STOP45 cells were induced with Dox and NaB for 48 h, and then fixed and stained with anti-pFilamin A antibody with Alexa-555 secondary antibody, and finally visualized with confocal fluorescence microscopy. The percentages of pFilamin A-positive cells in cells undergoing lytic reactivation were calculated in three independent experiments and shown. **, p < 0.01; t test. G. The iSLK.Bac16 cells were induced with Dox and NaB for 48 h, and then fixed and stained with anti-ORF45 antibody with Alexa-647 secondary antibody and anti-pFLNA antibody with Alexa-555 secondary antibody, and finally visualized with confocal fluorescence microscopy. The subcellular co-localization of ORF45 and pFilamin A were analyzed and shown.</p> |
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