Fig. 1: Na⁺-dependent energy depletion and killing of HCC cells treated with Monensin.

a Dose dependent analysis of the cytotoxic effects of 20 h treatments with Monensin (M) on C1C7, HepG2 cells and hepatocytes in DMEM ± Na⁺. b Effects of 20 h treatments with 10 μM Monensin (M) on the viability of C1C7, HepG2 cells and hepatocytes in DMEM ± Na⁺. c Representative in vivo cell living images of intracellular Na+ increase (Green: ING) and appearance of cell death (Violet: TO-PRO3) of C1C7 cell exposed to Monensin 10 μM in DMEM + Na⁺. Time frame 30′: T0 0′-T1 30′-T2 60′- T3 90′ -T4 120′. d The ATP levels (nmol/10⁶ cells) of Hp, C1C7 and HepG2 cells upon 8 h of 10 μM Monensin (M) treatments in DMEM ± Na⁺ (levels at T0: Hp = 17.5 ±  2.1; C1C7 = 19.9 ±  1.8;HepG2 18.9 ±  1.9). e Viability of Hp, C1C7 and HepG2 cells with 4 h treatment of 10 μM Monensin (M) in Krebs ± Na⁺ plus or minus glucose (glu) 5 mM. f Intracellular ATP levels of Hp, C1C7 and HepG2 cells with 90 min treatments of 10 μM Monensin (M) in Krebs ±  Na⁺ plus or minus glucose (glu) 5 mM. Results are expressed as % of controls (n = 8 independent experiments). Symbols represent the average while bars represent the standard deviation. Green lines represent the presence of sodium in the extracellular medium. ***P < 0.001 with O-way Analysis of Variance (ANOVA) Bonferroni Multiple Comparisons Test. C1C7 + M + Na⁺ and HepG2 + M + Na⁺ significantly different from C1C7 + M + Na⁺ and HepG2 + M + Na⁺; Hepatocytes +M ± Na⁺ in all panels; and C1C7 + M + Na⁺ + glu and HepG2 + M + Na⁺ + glu in panel e (except the 2nd hour) and f.