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. 2022 May 19;5:483. doi: 10.1038/s42003-022-03427-4

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a, b Immunofluorescence of Flag-STAT1 in HeLa cells transfected with AcGFP, AcGFP-ORF6 WT, or AcGFP-ORF6Δ9 under an untreated condition a or following IFN- γ stimulation b. Anti-GFP and anti-Flag antibodies were used for the detection of AcGFP (green) and Flag-STAT1 (red), respectively. DAPI was used to stain the DNA (blue). Scale bars: 30 μm. c, d The graphs represent the relative fluorescence values of Flag-STAT1 in the nucleus compared to those of the whole cells under an untreated condition in a or following IFN- γ stimulation in b, respectively. Signal intensities from total 45 nuclei from two independent experiments. ***P < 0.001, one-way ANOVA. error bars represent SD. e Immunoprecipitation (IP) of Flag-STAT1 from HEK293 cells transfected with AcGFP-ORF6 which is tagged the HA sequence following stimulation with IFN-γ (+IFN). Normal mouse IgG was used as a negative control for IP. Input was 1/130 dilution of cell lysates used for the reaction. Flag-STAT1 and AcGFP-ORF6 were detected by either anti-Flag or anti-HA antibodies, respectively (WB). The stimulation of IFN-γ was identified by the detection of PY-STAT1 using anti-PY-STAT1 antibody. Light chain indicates the precipitated antibodies.