Figure 14.

Effect of human ankyrin repeat and suppressor of cytokine signalling (SOCS) box protein 9 (ASB9) on creatine kinase (CK) activity. (a) The cellular CK activity was measured during a 4-day culture of the human embryonic kidney 293 (HEK293) cells that stably express null, Myc-tagged ASB9, or ASB9ΔSOCS. The cell lysates (total cells), cytosolic fraction and mitochondrial fraction were prepared, and the CK activity was determined. (b) The cell lysates (total cells), cytosolic fraction and mitochondrial fraction of human peripheral blood mononuclear cells (hPBMCs), normal hepatocytes and human cell lines were prepared, and the CK activity was determined. (c) The immunocomplex proteins of the anti-CKB or anti-ubiquitous mitochondrial CK (uMtCK) antibodies were analysed by CK activity assay. Specific CK activity was defined in terms of units per cell (U/cell). The immunoprecipitated proteins were detected by immunoblotting analysis with an anti-CKB antibody and an anti-Myc antibody (for ASB9) (lower panel).