Fig. 4

Glutamine metabolism is altered in KL-11743 treated cells. A NB4, THP-1, and MOLT-4 were treated with DMSO or KL-11743 (500 nM) for 24 h, followed by YSI media analysis of glutamine and glutamate concentrations. Average rates are graphed ± SEM, with negative values indicating consumption and positive values indicating secretion. B NB4, THP-1, and MOLT-4 were cultured in complete media (( +)Q) or glutamine-free ((-)Q) media treated with DMSO or KL-11743 (500 nM) for 24 h. Viability was determined by CellTiter Glo kit, and values were normalized with DMSO-treated samples defined as 100% viability and samples co-treated with cycloheximide (50 μg/mL) and ABT-737 (1 μM) as 0% viability (data not shown). C–E NB4, THP-1, and MOLT-4 were treated with DMSO or KL-11743 (500 nM) for 18 h prior to changing them into glutamine-free ((-)Q) media for 6 h. OCR was measured using an Agilent XF Cell Mito Stress Test. O: oligomycin (1 μM), F: FCCP (1 μM), R + A: rotenone (0.5 μM) + antimycin A (0.5 μM). F Quantification of basal and maximal respiration from C, calculated as the average of the 3 reads preceding injection of oligomycin (O), and as the average of the 3 reads immediately following infection of FCCP (F), respectively. G Quantification of basal and maximal respiration from D, calculated as the average of the 3 reads preceding injection of oligomycin (O), and as the average of the 3 reads immediately following infection of FCCP (F), respectively. H Quantification of basal and maximal respiration from E, calculated as the average of the 3 reads preceding injection of oligomycin (O), and as the average of the 3 reads immediately following infection of FCCP (F), respectively. Panels A–B are presented as mean values ± SEM of 3 technical replicates; panels C–H are mean values ± SEM of at least 5 technical replicates. Individual dots in bar graphs represent technical replicates