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. 2019 Sep 9;11(9):1336. doi: 10.3390/cancers11091336

Figure 7.

Figure 7

Piper genus-derived compounds induce cell death and mitochondrial dysfunction. (A) Immunofluorescence (confocal) imaging of cleaved-caspase 7 (punctate red pattern) in U373 cells treated for 6 h in the absence (CTRL, control) and in the presence of gibbilimbol B/eriopodol A (30 µg/mL) or erioquinol (10 µg/mL). 4’,6-diamidine-2’-phenylindole dihydrochloride (DAPI) (blue) and phalloidin (green) were used for nuclei and cytoskeleton detection, respectively. Scale bar = 25 µm. (B) Bright field microscopy (upper panels) and DAPI staining (lower panels) of U373 cells treated for 6 h in the absence (CTRL) and in the presence of of gibbilimbol B/eriopodol A (30 µg/mL) or erioquinol (10 µg/mL). Scale bars = 50 µm (bright field) and 10 µm (DAPI). Lower panels represent enlarged image details. (C) Quantitative analysis of tetramethylrhodamine methyl ester (TMRM) fluorescence changes over time in MCF7 (left panel) and U373 (righ panel) cells in the absence (CTRL) and in the presence of gibbilimbol B/eriopodol A (30 µg/mL) or erioquinol (10 µg/mL). Results are expressed by setting TMRM fluorescence in the respective control (vehicle-treated) samples, i.e., absence of compounds, as 1. *** p < 0.0001 relative to CTRL. Images and data are representative of four independent experiments.