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  Indian J Med Microbiol
 

Figure 2: AMP-activated protein kinase (AMPK) activation is required for methanol extract from Impatiens balsamina (MEIB) induced apoptosis. HSC-2 cells were treated with dimethyl sulfoxide (DMSO) or 60 μg/ml of MEIB for 24 h. (a) Whole-cell lysates were analyzed by Western blot analysis using antibodies against phosphorylation of AMPK (p-AMPK), AMPK, phosphorylation of mammalian target of rapamycin (p-mTOR), mTOR, p-p70S6 kinase, p70S6 kinase, p-p38, p38, p-ERK and ERK. Actin was used as a loading control, (b) HSC-2 cells treated with 60 μg/ml of MEIB were harvested at different time points (0, 6, 12 or 24 h), and analyzed by Western blot analysis using antibodies against p-AMPK, AMPK, p-mTOR, mTOR, p-p70S6 kinase, p70S6 kinase, cleaved caspase-3 and cleaved poly(ADP-ribose) polymerase. (c) The apoptotic effect of MEIB was invesitgated by 4'-6-diamidino-2-phenylindole (DAPI) staining. Nuclear condensation and DNA fragmentation were initially observed by fluorescence microscopy (×400); subsequently, DAPI-stained cells were quantified. The numbers of apoptotic cells were expressed as the means ± standard deviation of triplicate experiments. *P < 0.05 compared with the DMSO-treated group

Figure 2: AMP-activated protein kinase (AMPK) activation is required for methanol extract from <i>Impatiens balsamina</i> (MEIB) induced apoptosis. HSC-2 cells were treated with dimethyl sulfoxide (DMSO) or 60 μg/ml of MEIB for 24 h. (a) Whole-cell lysates were analyzed by Western blot analysis using antibodies against phosphorylation of AMPK (p-AMPK), AMPK, phosphorylation of mammalian target of rapamycin (p-mTOR), mTOR, p-p70S6 kinase, p70S6 kinase, p-p38, p38, p-ERK and ERK. Actin was used as a loading control, (b) HSC-2 cells treated with 60 μg/ml of MEIB were harvested at different time points (0, 6, 12 or 24 h), and analyzed by Western blot analysis using antibodies against p-AMPK, AMPK, p-mTOR, mTOR, p-p70S6 kinase, p70S6 kinase, cleaved caspase-3 and cleaved poly(ADP-ribose) polymerase. (c) The apoptotic effect of MEIB was invesitgated by 4'-6-diamidino-2-phenylindole (DAPI) staining. Nuclear condensation and DNA fragmentation were initially observed by fluorescence microscopy (×400); subsequently, DAPI-stained cells were quantified. The numbers of apoptotic cells were expressed as the means ± standard deviation of triplicate experiments. *<i>P</i> < 0.05 compared with the DMSO-treated group