Nelfinavir Inhibits Intra-Mitochondrial Calcium Influx and Protects Brain against Hypoxic-Ischemic Injury in Neonatal Mice
Figure 5
Nelfinavir does not inhibit mitochondrial Ca2+ uniporter.
A – The experiment controlling mitochondrial specificity for TMRE and Rhodamine 123 (R123) fluoroprobes. Note a drastic decrease in Ψm fluorescence following FCCP (0.5 µM) supplementation. Scale bar = 10 µm. B – One of three highly reproducible tracings of mitochondrial Ca2+ buffering capacity in organelles pre-treated with RuR (1 µM), NLF (4.4 µM) or vehicle. Note, that only RuR completely inhibited Ca2+ up-take by mitochondria, while NLF, virtually, had no effect. C – One of the four highly reproducible tracings of changes in the safranin (Ψm) fluorescence in response to mitochondrial supplementation (indicated) and addition of 10 nmoles of Ca2+ pulses to mitochondria pre-incubated with RuR, NLF or vehicle. Note, only the RuR prevented the collapse of Ψm in response to Ca2+ challenge. Brain mitochondria were isolated from naïve p10 mice, substrate: succinate-glutamate (see also methods).