Fig. 4
Decreased ATP level and induction of autophagy in E260-treated SW620 cells. ATP levels in SW620 a and Hfb b cells treated with vehicle (blue histogram), or 2 µM E260 (gray histogram) for 12 h; n = 4, ±SE. c Protein lysates from SW620 cells treated with control or 2 µM E260 for 12 h were subjected to WB analysis of the total expression or phosphorylation level (p-X) of selected proteins from the AMPK–mTOR signaling pathway. d, e TEM analysis of SW620 cells treated with 2 µM E260 for 12 h. Red arrows indicate autophagosomes. N nucleus. Bars represent 2 µm in d and 500 nm in e. Immunostaining for LC3 (green) of SW620 cells treated with vehicles (f, h) or with 2 µM E260 (g, i) for 12 h. Nuclei were visualized with Hoechst (blue). Bars represent 10 µm. j Protein lysates from SW620 cells treated with control or 2 µM E260 for 12 or 20 h were subjected to WB analysis using anti-LC3 antibody. k ATP levels in SW620 cells treated for 12 h with vehicle (gray), 2 µM E260 (black), 2 µM E260, and 5 mM 3-MA (blue), and 5 mM 3-MA alone (red); n = 3, ±SE. l Cell death levels in SW620 cells treated for 16 h with vehicle (gray), 2 µM E260 (black), 2 µM E260, and 5 mM 3-MA (blue), or 5 mM 3-MA alone (red); n = 4, ±SE
