Isoliquiritigenin Induces Mitochondrial Dysfunction and Apoptosis by Inhibiting mitoNEET in a Reactive Oxygen Species-Dependent Manner in A375 Human Melanoma Cells

Figure 3

ISL triggers the production of reactive oxygen species, which contributes to ISL-induced apoptosis and mitochondrial dysfunction in A375 cells. (a) Representative confocal microscopy images of mitochondrial network in A375 cells treated with or without ISL. (b) Quantification of the MitoSOX Red and MitoTracker Green fluorescence intensity detected by flow cytometry. (c) Representative confocal microscopy images of DCFH-DA and quantification of the DCFH-DA fluorescence intensity by flow cytometry. (d) Analysis of the GSSH/GSH ratio. (e) The inhibition rates of A375 cells treated by ISL alone or ISL cotreated with NAC or BSO. (f) Phase-contrast micrographs (200x) showing morphological changes in A375 cells treated with ISL alone or ISL cotreated with NAC or BSO. (g) Apoptosis rates of A375 cells determined by annexin V/PI double staining in response to indicated treatments. (h) Representative confocal microscopy images of the JC-1 probe in A375 cells treated by ISL alone or ISL cotreated with NAC or BSO. (i) Quantification of the JC-1 fluorescence intensity in A375 cells by flow cytometry. (j) Cytosolic and mitochondrial cytochrome c concentrations in A375 cells treated by ISL alone or ISL cotreated with NAC or BSO. , , and versus control; versus ISL.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)