Fig. 1.

Identification of SNX27 as an OTULIN interactor. a Selective protein enrichment after OTULIN ABP-PD. Volcano plot illustrating the enrichment of proteins identified by LC-MS/MS after PD in the presence or absence of OTULIN ABP. Curves depict significant enrichment or depletion, respectively. b Selective depletion of proteins from OTULIN ABP by PR-619 treatment. Volcano plot demonstrates loss of protein binding in samples treated with PR-619 before OTULIN ABP incubation and PD. Curves depict significant enrichment or depletion, respectively. c OTULIN-ABP PD was performed under conditions used for LC-MS/MS and binding of HOIP and SNX27 was analyzed by WB. d OTULIN ABP-PD was performed from extracts of parental or OTULIN KO Jurkat T cells. Co-precipitation of HOIP and SNX27 was analyzed by WB. e OTULIN ABP-PD was performed from extracts of parental or HOIP KO Jurkat T cells. Co-precipitation of HOIP and SNX27 was analyzed by WB. f Enrichment of proteins by PD of SF-OTULIN. Volcano plot depicts significant enrichment of proteins after Strep-PD from extracts of OTULIN-deficient Jurkat T cells expressing SF-OTULIN or mock control. Curves depict significant enrichment or depletion, respectively. g OTULIN-SNX27 interaction in reconstituted OTULIN KO Jurkat T cells was analyzed by WB after Strep-PD. h OTULIN-IP and SNX27-IP from extracts of Jurkat T cells. Rabbit or mouse IgG antibodies were used as isotype controls and protein binding was analyzed by WB. i OTULIN-IP from extracts of WT and OTULIN KO MEFs. Co-immunoprecipitation of SNX27 was analyzed by WB. j ABP-PD (left) and OTULIN-IP (right) from murine primary CD4 T cell extracts. Rabbit IgG antibody was used as an isotype control. Interaction of SNX27 to murine OTULIN-ABP complex and OTULIN was assessed by WB. Source data are provided as a Source Data file