Fig. 4

As characteristic of certain E3 ubiquitin ligases, NSPA becomes ubiquitinated in a cell-based ubiquitination assay. a Schematic representation of Cullin9/Parc and NSPA indicating linear domain organization. b NSPA ZZ-type finger domains (ZZ1 and ZZ2) sequence alignment with BRcat domain of Cullin9/Parc E3 ubiquitin ligase, conserved cysteine, and histidine residues are colored. c HEK293 cells transfected with NSPA-mCherry and myc-6xHis-ubiquitin and subjected to denaturing Ni-NTA pulldown followed by immunoblot with anti-mCherry antibodies clearly show NSPA ubiquitination. Lanes 1–2, 3–4, and 5–6 are duplicates of the same experiment