Figure 1 (IMAGE)
Caption
Inhibitory effect of CADD522 on mitochondrial OCR. (A) (A) Acute OCR changes were determined in T47D-RUNX2 and -Empty cells with no prior CADD522 treatment. Cells seeded in Seahorse plates were incubated overnight in growth medium without G418, and then replenished with Seahorse-Certified Medium on the assay day. Vehicle control (0.1% DMSO), Oligomycin or CADD522 were directly injected into port A of the XF Extracellular Flux Analyzer, and OCR was measured. Data (% OCR) are presented as the percentage of the basal OCR value at t = 12 min (100%). Experiments were performed in four replicates and repeated twice (mean ± SD). O, Oligomycin (1 μg/ml); F, FCCP (1 μM, port B); P, pyruvate (10 mM, port C); AA, antimycin A (1 μM, port D). Red line, maximal OCR value of the Control at t = 57 min. (B) Cells were treated with CADD522 for 6~72 hr and replenished with Seahorse-Certified Medium with no CADD522. OCR was measured in the XF Extracellular Flux Analyzer. Data (mean ± SD) are presented as OCR (pMoles/min/μg protein). (C) Mitochondrial respiratory parameters were calculated as described in Material and Methods: Maximal respiratory capacity (MRC), mitochondrial reserve capacity (RC), ATP production-linked respiration (AP), proton leak-linked respiration (PL), baseline respiration (BL), and non-mitochondrial respiration (NM). Experiments were performed in four replicates and data are expressed as mean ± SE from two independent experiments. *P < 0.05 compared to Empty cells with vehicle control (0.1% DMSO); #P < 0.05 compared to RUNX2-expressing cells with vehicle control.
Credit
Correspondence to - Myoung Sook Kim - mkim@som.umaryland.edu and Antonino Passaniti - TPassaniti@som.umaryland.edu
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Copyright: © 2020 Kim et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
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