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. Author manuscript; available in PMC: 2017 Aug 1.
Published in final edited form as: J Mol Cell Cardiol. 2016 Jun 1;97:191–196. doi: 10.1016/j.yjmcc.2016.05.015

Figure 2. Effects of MTP-131 on primary myotube and HUVEC cellular respiratory function.

Figure 2

Cellular respiration was assessed using the Seahorse XF24 analyzer in both control (normoxia with normal culture media) and HND (hypoxia with HBSS) muscle and endothelial cells. A. Primary BALB/c muscle cells (vehicle vs. MTP-131) were exposed to 3-hr HND, after which cellular respiration was assessed immediately following using a sequential protocol with additions of 1μg/ml oligomycin (ATP synthase inhibitor), 1μM FCCP (chemical uncoupler), and 2.5μM antimycin A (mitochondrial complex III inhibitor). B. Vehicle-treated primary muscle cells and MTP-131 primary muscle cells were subjected to 3-hr HND and allowed to recover in normal media for 24 hours before assaying cell respiration. C. Basal respiration rates (prior to addition of oligomycin) expressed as a percentage of the control (normoxia) cells with no recovery (from panel A) and 24h recovery (from panel B). D. Maximal respiratory capacity (addition of FCCP) expressed as a percentage of the control (normoxia) cells with no recovery (from panel A) and 24h recovery (from panel B). E. BALB/c primary muscle cells were treated with MTP-131 and immediately subjected to 3-hr HND (no pre-treatment) and allowed to recovery 24 hours before cell respiration assay. F. BALB/c primary muscle cells were subjected to 3-hr HND and then treated with MTP-131 after HND (post-treatment) and allowed to recovery 24 hours before cell respiration assay. G. Basal respiration rates (prior to addition of oligomycin) expressed as a percentage of the control (normoxia) cells (from panels E and F) and 24h recovery. H. Maximal respiratory capacity (addition of FCCP) expressed as a percentage of the control (normoxia) cells (from panels E and F). I. Human endothelial cells (vehicle vs. MTP-131) were exposed to 3-hr HND, after which cellular respiration was assessed immediately following using a sequential protocol with additions of 1μg/ml oligomycin (ATP synthase inhibitor), 1μM FCCP (chemical uncoupler), and 2.5μM antimycin A (mitochondrial complex III inhibitor). J. Vehicle-treated HUVECs and MTP-131 treated HUVECs were subjected to 3-hr HND and allowed to recover in normal media for 24 hours before assaying cell respiration. K. Basal respiration rates (prior to addition of oligomycin) expressed as a percentage of the control (normoxia) cells with no recovery (from panel E) and 24h recovery (from panel F). L. Maximal respiratory capacity (addition of FCCP) expressed as a percentage of the control (normoxia) cells with no recovery (from panel E) and 24h recovery (from panel F). a P<0.05 vs. no recovery (time effect). bP<0.05 vs. vehicle-treated control (drug effect).