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. 2020 May 29;117(24):13740–13749. doi: 10.1073/pnas.1922884117

Fig. 4.

Fig. 4.

HBZ modifies the IL-10/JAK/STAT signaling pathway. (A) Fluorescence dilution assay of CD4+ T cells from WT or HBZ-Tg mice. Splenic CD4+ T cells were labeled with 5 μM CellTrace Violet and stimulated by anti-CD3 antibodies with or without IL-10. At 48 h after stimulation, CellTrace Violet was measured by flow cytometry (two-way ANOVA with Turkey’s multiple comparisons). (B) Coimmunoprecipitation of HBZ and human STAT1 or STAT3. The indicated expression vectors were cotransfected into HEK293T cells, and protein interactions were analyzed by immunoprecipitation. (C) Interaction of HBZ mutants with STAT1 or STAT3 was analyzed by immunoprecipitation. (D and E) Interaction between HBZ and STAT proteins in ATL-43T(−). A protein extract of ATL-43T(−) cells was subjected to immunoprecipitation with anti-HBZ antibody or control IgG, and STAT proteins were detected by anti-STAT1, anti-STAT3, anti-STAT5, or anti-STAT6 antibody. (F) Colocalization of HBZ and STAT proteins. HBZ-myc and STAT1 or STAT3-3xFLAG were transfected into Jurkat cells by electroporation. Staining was performed using antibodies against myc (green) and FLAG (red). Nuclei were stained with DAPI (blue).