Fig. 9

The UBAN domain of OPTN and the dead domain of RIPK1 mediate their interaction. A After 48 h of simultaneous overexpression of GFP-OPTN and mCherry-RIPK1 in BV2 cells, cells on the slide were fixed in PFA and stained with DAPI, followed by imaging using confocal microscopy. B The protein was precipitated using an OPTN antibody in BV2 cells and then detected using an RIPK1 antibody. C In HEK293T cells, both HA-OPTN and Flag-RIPK1 were overexpressed, which were then precipitated using an anti-Flag antibody and detected using an HA antibody. D Structural diagram of the OPTN protein. E Schematic diagram of RIPK1’s protein structure. F Full-length HA-OPTN, NEMO-truncated OPTN or UBAN-truncated OPTN were ectopically expressed in Flag-RIPK1-transfected HEK293T cells. Then, the proteins were immunoprecipitated using an anti-Flag antibody, followed by detection with an anti-HA antibody. G Full-length RIPK1, RIPK1 with a truncated protein-like kinase domain and RIPK1 with a truncated dead domain were ectopically expressed in HA-OPTN-transfected 293 T cells. Then, the proteins were immunoprecipitated using an anti-HA antibody, followed by detection with an anti-Flag antibody