Figure 4.

Assessment of mitochondrial dysfunction and release of Cyt-C. (a–d) LNCaP cells and normal HSG salivary cells overexpressing Nek1 or T141A mutant were monitored for OCR and ECAR with a Seahorse analyzer according to the manufacturer instructions. Cells in A-C were sequentially treated with oligomycin, FCCP, and Antymycin A/Rotenone. Cells in D were sequentially treated with Glucose after 3 h starvation, oligomycin, and finally 2-deoxyglucose (2-DG). (e) Expression of wt or T141A mutant Nek1 in HSG cells, since these cells were not previously characterized. (f–i) Western blots of Cyt-C in cytoplasmic and mitochondrial fractions of Nek1 and t14A mutant overexpressing cells with and without treatment with doxorubicin (200 nM) for 12 h.