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. 2014 Oct 1;33(10):652–666. doi: 10.1089/dna.2014.2366

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Copyright 2014, Mary Ann Liebert, Inc.

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FIG. 2. — Tazarotene triggered apoptotic events in BCC cells. (A) BCC cells were stained with JC-1 fluorescent dye, and mitochondrial membrane potential (Δψm) was analyzed by flow cytometry. The means±SD of the experimental triplicates are presented in the bar graph at the bottom. (B) The cells were treated with indicated concentrations of tazarotene for 24 h. Cytosolic cell lysates were prepared, resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and immunoblotted with the indicated antibodies to detect cytochrome c. Cytochrome c oxidase subunit IV (COX IV) served as a mitochondrial marker. (C) Total cell lysates were prepared to detect the non-cleaved and cleaved forms of caspase-9, caspase-3, and PARP. (D) The bands were analyzed by ImageJ (National Institute of Health) and normalized to actin. The means±SD of the three independent experiments are presented in the bar graph. (E) The effect of caspase-3 and caspase-9 inhibitors on 100 μM tazarotene-induced cell viability. The data represent the mean±SD from three wells. The data are representative of three independent experiments with similar results. (F) Total cell lysates were prepared and subjected to SDS-PAGE followed by Western blotting with antibodies to detect the expression of Bcl-2, Bcl-xl, Bax, Bak, XIAP, and Survivin. (G) The bands were analyzed by ImageJ and normalized to actin. The means±SD of the three independent experiments are presented in the bar graph. ^nsp>0.05, *p<0.05, **p<0.01, and ***p<0.001, compared with the 0.1% DMSO-treated group. ^#p<0.05, ^###p<0.001 compared with the 100 μM tazarotene-treated group.

FIG. 2. — Tazarotene triggered apoptotic events in BCC cells. (A) BCC cells were stained with JC-1 fluorescent dye, and mitochondrial membrane potential (Δψm) was analyzed by flow cytometry. The means±SD of the experimental triplicates are presented in the bar graph at the bottom. (B) The cells were treated with indicated concentrations of tazarotene for 24 h. Cytosolic cell lysates were prepared, resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and immunoblotted with the indicated antibodies to detect cytochrome c. Cytochrome c oxidase subunit IV (COX IV) served as a mitochondrial marker. (C) Total cell lysates were prepared to detect the non-cleaved and cleaved forms of caspase-9, caspase-3, and PARP. (D) The bands were analyzed by ImageJ (National Institute of Health) and normalized to actin. The means±SD of the three independent experiments are presented in the bar graph. (E) The effect of caspase-3 and caspase-9 inhibitors on 100 μM tazarotene-induced cell viability. The data represent the mean±SD from three wells. The data are representative of three independent experiments with similar results. (F) Total cell lysates were prepared and subjected to SDS-PAGE followed by Western blotting with antibodies to detect the expression of Bcl-2, Bcl-xl, Bax, Bak, XIAP, and Survivin. (G) The bands were analyzed by ImageJ and normalized to actin. The means±SD of the three independent experiments are presented in the bar graph. ^nsp>0.05, *p<0.05, **p<0.01, and ***p<0.001, compared with the 0.1% DMSO-treated group. ^#p<0.05, ^###p<0.001 compared with the 100 μM tazarotene-treated group.