Fig. 3: KCTD7–Cullin-3 ubiquitin ligase regulates calpain activity via atypical ubiquitination. | Cell Discovery

Fig. 3: KCTD7–Cullin-3 ubiquitin ligase regulates calpain activity via atypical ubiquitination.

From: Calpain activity is negatively regulated by a KCTD7–Cullin-3 complex via non-degradative ubiquitination

Fig. 3

a In vivo ubiquitination assay for calpain 1, calpain 2, and CAPNS1 performed by coexpressing either protein with HA-Ub in HEK293 cells for 36 h. b In vivo ubiquitination assay in HEK293 cells for calpain 1, calpain 2, and CAPNS1 using the indicated ubiquitin mutants. HA-Ub K0 has all lysine residues mutated. c In vivo ubiquitination assay for calpain 1, calpain 2, and CAPNS1 performed by expressing Cullin-3 and KCTD7 alone or in combination in HEK293 cells for 36 h. d In vivo ubiquitination assay for calpain 1, calpain 2, and CAPNS1 performed upon simultaneous expression of KCTD7 and knock-down of Cullin-3 in HEK293 cells for 48 h. e In vivo ubiquitination assay in HEK293 cells for calpain 1, calpain 2, and CAPNS1 performed by expressing the indicated ubiquitin mutants alone or in combination with KCTD7.

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