Extended Data Fig. 4 ∣. Regnase paralogs cannot complement for Roquin loss of function.

(a) Workflow of in vitro reconstitution experiments. (b) Mutations introduced in Regnase-1 coding sequence (marked in red) for the generation of an antibody-invisible GFP-tagged Regnase-1 construct (GFP-Regnase-1^invis). (c) Flow cytometry analysis of Regnase-1 expression after retroviral transduction of WT CD4⁺ T cells with GFP-Regnase-1 or GFP-Regnase-1^invis constructs. (d) WT or iDKO CD4⁺ T cells were retrovirally transduced with GFP–Regnase-1 or GFP-Regnase-1^invis. Contour plots of flow cytometry analysis of CTLA-4 expression in dependence of the GFP expression level. (e, f) Flow cytometry analysis of ICOS or Regnase-1 expression after retroviral transduction of iKO or iDKO CD4⁺ T cells with the indicated GFP-fusion protein. Contour plots of histograms shown in Fig. 4c. (d–f) Data are representative of n = 3 independent experiments.