262 389 163 310 ADP titration Seahorse XF24 Mitochondrial integrity test Seahorse XF24 b d ADP titration Clarktype electrode 500 701 1000 403 511 1027 a Mitochondrial integrity test Clarktype electrode ID: 630571
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Slide1
Supplemental Figure
2
~2.03
~2.62
~3.89
~1.63
~3.10
ADP titration (Seahorse XF24)
Mitochondrial integrity test (Seahorse XF24)
b
d
ADP titration (Clark-type electrode)
~5.00
~7.01
~10.00
~4.03
~5.11
~10.27
a
Mitochondrial integrity test (Clark-type electrode)
cSlide2
Figure S2: Validation of Seahorse XF24 method for measuring respiration in isolated mitochondria. A) Assessment of respiratory control ratio (RCR) by Clark-type electrode in muscle (left panel) liver (right panel) mitochondria. Following measurement of state 2, ADP was sequentially injected to a final concentration of 0.5 mM to test state 3 respiration rates.
Antimycin A (Ant A; 10 μM) was then injected to inhibit respiration.
B) Assessment of RCR by Seahorse XF24 in muscle (left panel) liver (right panel) mitochondria. Following measurement of state 2, ADP was sequentially injected to a final concentration of 0.5 mM. Antimycin A (Ant A; 4 μM) was then injected. RCR was estimated as the ratio of state 3/state 2. n=2, mean±SEM
. C) Assessment of mitochondrial integrity by Clark-type electrode in muscle (left panel) liver (right panel)
mitochondria. Following measurement of state 2, state 3 respiration was stimulated using ADP (final concentration of 0.05 mM) followed by treatment with cytochrome C (final concentration of
10 μM). Antimycin A (Ant A; 10
μM) was then injected to deactivate respiration. D) Assessment of mitochondrial integrity by Seahorse XF24 in muscle (left panel)
and liver (right panel) mitochondria. Following measurement of state 2, state 3 respiration was stimulated using ADP (final concentration of
0.1 mM) followed by treatment with sequential treatment with cytochrome C (final concentration of 10 μM
). Antimycin A (Ant A; 4 μM) was then injected. n=2, mean±SEM
.