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Further confirmation that mitochondrial dysfunction contributes to OXY activity is provided by microscopic analysis using the fluorescent indicator JC-1 which aggregates in healthy mitochondria and emits a red fluorescence.
The intracellular oxidant status of freshly isolated tracheal cells from Cftr−/− mice and their Cftr+/+ littermates was analyzed using the fluorescent indicator CM-H2DCFDA.
The mitochondrial membrane potential was monitored using the fluorescent indicator JC-1.
ROS were detected using the fluorescent indicator dye DCFH2-DA (Calbiochem).
[Ca2+]i was measured using the fluorescent indicator, fura-2, as previously described [ 35].
A possible role for dP32 in maintaining ΔΨm was examined using the fluorescent indicator tetramethylrhodamine ethyl ester (TMRE).
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[Ca2+]i was measured in HEK cells transfected with CaSR-EGFP or mGluR1 using the fluorescent indicators, X-rhod-1 or Fluo-4 respectively.
A second experiment used the fluorescent indicator JC-1 to measure changes in mitochondrial membrane potential (the difference in electrical potential between the inside and outside of the mitochondrial membrane) following exposure to Zn2+.
Methods: In this paper, we describe the development of a microplate reader assay for the assessment of [Ca2+]i in a primary culture of rat hippocampal neurons maintained in Neurobasal medium using the fluorescent calcium indicator, fluo-3.
We measured changes in [Ca2+]i using the fluorescent Ca2+ indicator Fluo-4 (Molecular Probes, Eugene, OR).
Changes in intracellular calcium were determined using the fluorescent calcium indicator Fluo-4 AM (Invitrogen).
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