Figure 8.

Knockdown of UQCRB leads to a decrease in mitochondrial membrane potential, oxidative mitochondrial metabolism, and IFN-γ production. NK-92 cells were transduced with an empty vector or a pLKO.1 vector containing UQCRB-specific shRNA. (a) Western blot of UQCRB and β-actin in the control and shUQCRB NK-92 lines (left) and quantification by densitometry (right). (b) Representative FACS plots (left) and cumulative fold differences in MitoTracker MFI values (right) from each indicated NK-92 cell line. Experiments were replicated twice. (c) OCR profiles of the control and shARID5B NK-92 cell lines in a representative experiment and averages for maximal respiration, ATP-linked respiration, and SRC. (d) ECAR profiles for the indicated NK-92 cell lines in a representative experiment and averages for maximal glycolysis and glycolytic reserve. All cumulative OCR and ECAR results were replicated twice. (e) Quantification of ATP in control and shUQCRB cell lines. (f) Control and shUQCRB NK-92 cells were cultured overnight with and without IL-12 and IL-18 before FACS analysis of NK cell functional readouts. Shown are representative FACS plots and cumulative data of the frequencies of IFN-γ expression by NK-92 cells. (e and f) Experiments were replicated twice. Error bars represent SEM. Paired Student’s t tests were used to determine statistical significance. *P < 0.05. n.s., not significant.