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Figure 4 | Molecular Cancer

Figure 4

From: A combination of indole-3-carbinol and genistein synergistically induces apoptosis in human colon cancer HT-29 cells by inhibiting Akt phosphorylation and progression of autophagy

Figure 4

Detection of autophagosomes following co-treatment. A, After exposure to a combination of I3C (300 μmol/L) and genistein (40 μmol/L) for the periods indicated, cell lysates were subjected to western blotting with an anti-LC3 antibody. β-actin was used as a loading control. B, After exposure to the indicated agents for 48 h, cell lysates were subjected to western blotting with an anti-LC3 antibody. β-actin was used as a loading control. -, treated with DMSO: C, HT-29 cells were exposed to DMSO (control) or a combination of I3C (300 μmol/L) and genistein (40 μmol/L) for 12 h with or without 10 mmol/L of 3-methyladenine (3-MA). As a positive control for the induction of autophagy, cells were subjected to amino acid starvation (AAS) by incubating them in amino acid-deprived medium for 12 h. After the incubation, cells were subjected to immunofluorescent staining for LC3 as described in Materials and Methods. Bars, 20 μm. D, HT-29 cells were exposed to DMSO (control) or a combination of I3C (300 μmol/L) and genistein (40 μmol/L) in complete medium or subjected to AAS for 12 h. The cells were then harvested and subjected to transmission electron microscopy as described in Materials and Methods. White arrows, autophagic vesicle. N, nucleus. GEN, genistein.

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