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Fig. 4 | Cell Communication and Signaling

Fig. 4

From: Deubiquitinase USP5 regulates RIPK1 driven pyroptosis in response to myocardial ischemic reperfusion injury

Fig. 4

USP5 reverses K63-linked ubiquitination of RIPK1. (A) Immunoblot analysis of lysates from HEK293T cells transfected with HA-tagged ubiquitin (HA-Ub), Flag-RIPK1and USP5 WT or C335A, followed by IP with anti-flag, probed with anti-HA or K63-Ub. (B) Immunoblot analysis of lysates from HEK293T cells transfected with HA-tagged K48-linked ubiquitin (K48-Ub) or HA-tagged K63-linked ubiquitin (K63-Ub), Flag-RIPK1 and Myc-USP5, followed by IP with anti-Myc, probed with anti-HA. (C) Immunoblot analysis of lysates from USP5+/+/USP5+/-mouse CMECs, followed by IP with anti-RIPK1, probed with anti-Ub, K48-Ub or K63-Ub. (D) Immunoblot analysis of extracts from HEK293T cells transfected with Flag-RIPK1and Myc-tagged USP5 or its mutants. (E) Immunoblot analysis of extracts from HCMECs transfected with USP5 wild type (WT), USP5 C335A or mutants of USP5. Similar results were obtained in three independent experiments

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Cell Communication and Signaling

ISSN: 1478-811X