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. 2019 Dec 5;159(3):309–321. doi: 10.1111/imm.13147

Figure 4.

Figure 4

USP7 regulates interferon type I (IFN‐I) ‐induced Janus kinase–signal transducer and activator of transcription (JAK–STAT1) activation through SOCS1. (a) Western blot analysis of STAT1 phosphorylation on tyrosine 701 (p‐STAT1) in HEK293T cells transfected with control shRNA (−) or three shUSP7 and then treated with IFN‐α (1000 IU/ml) for 30 min. (b) Western blot analysis of p‐STAT1 in HEK293T cells transfected with shCON or shUSP7 and then treated with IFN‐α (1000 IU/ml) as indicated. (c) Western blot analysis of p‐STAT1 in HEK293T cells transfected with empty vectors (CON) or Flag‐USP7 and then treated with IFN‐α (1000 IU/ml) as indicated. (d) Western blot analysis of p‐STAT1 in HEK293T cells transfected with Flag‐USP7 and then treated with IFN‐α (0, 1000, 3000 IU/ml) for 30 min. (e) Western blot analysis of p‐STAT1 in HEK293T cells transfected with empty vectors (−) or Flag‐USP7 (WT or inactive C223S mutant) and then treated with IFN‐α (1000 IU/ml) as indicated. (f) Western blot analysis of phosphorylated Tyk2 on tyrosine 1054/1055 (p‐Tyk2) and phosphorylated JAK1 on serine 1022/1023 (p‐JAK1) in HEK293T cells transfected with Flag‐USP7 and then treated with IFN‐α (1000 IU/ml) as indicated. (g) Western blot analysis of endogenous SOCS1 protein levels in stable SOCS1‐knockdown HeLa cells (shSOCS1: 1# and 2#) or stable control‐shRNA HeLa cells (shCON). (h) Western blot analysis of p‐STAT1 levels in stable SOCS1‐knockdown HeLa cells transfected with Flag‐USP7 and then treated with IFN‐α (1000 IU/ml) for 30 min