Reduced mitochondrial respiratory activities in CML CD34⁺CD38⁻ cells 24 hours after CXCL14 stimulation. (A) Experimental setup. BM cells from patients with primary CML were first stimulated with recombinant CXCL14 proteins or phosphate buffered saline (PBS) for 24 hours and collected for the detection of mitochondrial mass, ROS, mTOR, and OXPHOS pathway by FACS. (B) FACS profile showing gating strategy for CML CD34⁺CD38^– cells 24 hours after CXCL14 stimulation. (C) Representative histograms showing expression of CYC1, ATP2A2, phospho mTOR, OPA3, and SLC25A26. (D) The mean fluorescent intensities (MFI) of CYC1, ATP2A2, phospho mTOR, OPA3, and SLC25A26 in the CD34⁺CD38^– BM cells. Each dot represents data from 1 patient with CML. The numbers in the panel are P values determined by paired t test. (E) Histograms showing mitochondrial mass and ROS intensity in the CML CD34⁺CD38^– cells. (F) MFI of mitotracker red and ROS staining in the CD34⁺CD38^– BM cells. Each dot represents data from 1 patient with CML. The numbers in the panel are P values determined by paired t test. (G) Experimental setup for monitoring MMP by tetramethylrhodamine staining in K562 CML cells 24 hours after stimulation with CXCL14. The cells stimulated by PBS or CXCL14 were separated by CellTrace violet, and their MMPs were recorded continuously before and after the additions of complex I stimulus malate and pyruvate, and subsequently, OXPHOS uncoupler carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP). (H) Representative FACS profile showing dynamics of MMP (tetramethylrhodamine intensity) in the K562 cells after stimulation with PBS or CXCL14. The arrows indicate the addition of the corresponding reagents. See also supplemental Figure 9.