Figure 6.
Dex induces alterations in mitochondrial membrane potential (ΔΨm) and superoxide ( \(\mathbf{O}_{2}^{-}\)
) generation in AS-Hsp27-transfected MM.1R cells. (A) MM.1R cells were transiently transfected with either GFP-tagged AS-Hsp27 (WT) or with empty vector. Following transfections, GFP+ cells were selected by flow cytometry; treated with Dex (5 μM) for 24 hours, incubated with CMXRos for the last 20 minutes, and analyzed by flow cytometry to assay for alterations in ΔΨm. Results are mean ± SD of 3 independent experiments (P < .003). (B) AS-Hsp-27 or vector-transfected MM.1R cells were treated with Dex (5 μM) for 24 hours, harvested, stained with membrane permeable dye dihydroethidium (HE) for the last 15 minutes, and analyzed by flow cytometry. Results are mean ± SD of 3 independent experiments (P < .005). Superoxide anions oxidize HE to fluorescent ethidium, permitting analysis by flow cytometry (The Vantage, FACScan, Becton Dickinson) using excitation at 480 nm and emission at 630 nm.