Supplementary Figure 7: K27-linked polyubiquitination mediated the interaction among Rhbdd3, A20 and NEMO. | Nature Immunology

Supplementary Figure 7: K27-linked polyubiquitination mediated the interaction among Rhbdd3, A20 and NEMO.

From: Rhbdd3 controls autoimmunity by suppressing the production of IL-6 by dendritic cells via K27-linked ubiquitination of the regulator NEMO

Supplementary Figure 7

(a) Immunoblot of cell lysates immunoprecipitated by single or sequential V5 IP in HEK293 cells overexpressed with V5-tagged Rhbdd3 and HA-tagged one-K ubiquitin mutants. In single V5 IP, cell lysates were immunoprecipitated with anti-V5; in sequential V5 IP, proteins immunoprecipitated by anti-V5 were boiled in presence of 1% SDS and subjected to a second IP by anti-V5. (b) Immunoblot of cell lysates immunoprecipitated by Myc IP in HEK293 cells overexpressed with Flag-tagged Rhbdd3 and Myc-tagged A20. (c) Schematic of IP procedure in Fig. 7f. (d) Immunoblot of cell lysates immunoprecipitated by Myc IP followed by HA IP in HEK293 cells transfected as indicated. Data are from one representative of three independent experiments with similar results.

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