Fig. 2: Exposure of MC-3129 results in the dephosphorylation and mitochondrial translocation of cofilin. | Cell Death & Disease

Fig. 2: Exposure of MC-3129 results in the dephosphorylation and mitochondrial translocation of cofilin.

From: The cyclohexene derivative MC-3129 exhibits antileukemic activity via RhoA/ROCK1/PTEN/PI3K/Akt pathway-mediated mitochondrial translocation of cofilin

Fig. 2

U937 cells were treated with various MC-3129 concentrations for 24 h or with 10 μM of MC-3129 for different time intervals, as indicated. a Whole cell lysates were prepared and subjected to western blot analysis using antibodies against Mcl-1, phospho-Bad (p-Bad), Bad, Bcl-2, Bcl-xL, and GADPH. The cytosolic (Cytosol) and mitochondrial (Mito) fractions were also prepared and subjected to western blot analysis using antibodies against Bax, GADPH, and Cox IV. b Whole cell lysates, cytosolic (Cytosol), and mitochondrial (Mito) fractions were analyzed by western blot assay using antibodies against phospho-cofilin (p-cofilin), cofilin, GADPH, and Cox IV. U937 cells were transfected with control empty vectors or pseudophosphorylated (inactive, S3E) mutant plasmids or human cofilin dephosphorylated (active, S3A) mutant plasmids for 48 h, and then were treatment with 10 μM MC-3129 for 24 h. c, d Whole cell lysates and mitochondrial fractions were determined by immunoblotting. e The cell apoptosis was determined by flow cytometry using Annexin V/PI staining, the percentage of apoptotic cells was analyzed for three separate experiments (mean ± SD, **P < 0.01)

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