Figure 3

Regulation of mitochondrial function under hypoxic stress in DLBCL. (A–E) Representative oxygen consumption rate (OCR) traces from OxPhos subtype DLBCL cells. Cells were exposed to normoxia and hypoxia overnight and the next day mitochondrial respiration was measured during the sequential addition of uncoupler DNP plus 10 mM pyruvate, DNP alone twice, and finally an inhibitor of respiration antimycin A. Data are derived from n = 3 passages per cell line. (F and G) Maximal and basal respiration rates under hypoxia compared to control (cells maintained in normoxia). For maximal respiration *p values obtained for DLBCL cell lines were p < 0.028 for K422, p < 0.017 for Toledo, p < 0.001 for HLY, U2932 and Farage respectively. For basal respiration *p values obtained were p < 0.003 for K422, p < 0.040 for Toledo, p < 0.001 for HLY, U2932 and Farage respectively. (H) Cells were treated with CCCP overnight followed by treatment for 3 h before analysis with HCQ (to block lysosomal degradation). Flow cytometry was used to determine Mito Tracker green (MTG) staining, p < 0.0003 for difference in MTG intensity compared under normal and hypoxic culture conditions.