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. 2022 Nov 17;24(1):e55387. doi: 10.15252/embr.202255387

Figure EV5. KLHL22 interacts with IRF7 and promotes its ubiquitination.

Figure EV5

  1. Purification of recombinant proteins was performed by Coomassie blue staining and IB.
  2. CUL3‐KLHL22 E3 ligase catalyzed IRF7 ubiquitination in a cell‐free system. Ubiquitin and recombinant FLAG‐IRF7 were incubated with recombinant CUL3–RBX1 and KLHL22 or KLHL22‐Δ6xKelch.
  3. The associations between KLHL22 and various truncations of IRF7 were detected through the indicated IP and IB analyses.
  4. HEK293T cells were transfected with KLHL22 and IRF7 variants. IB of FLAG was performed followed by IP with an anti‐HA antibody.
  5. HEK293T cells were transfected with KLHL22, FLAG‐ubiquitin, and various IRF7 point mutants. IB of FLAG was performed, followed by IP with an anti‐HA antibody. The data are representative of at least three biologically independent experiments.

Data information: Please see Appendix Fig S8 for information regarding replicates, quantification, and statistical evaluation for biochemical data in this figure.

Source data are available online for this figure.