Cytotoxic Effects of Far-C on HepG2 Cells

The cytotoxic effects of Far-C against HepG2 cells was estimated using MTT assay, described previously [30 (link)]. Approximately, 5 × 10³ HepG2 cells were allowed to adhere in each well of a 96-well plate under standard culture condition. Post-adherence, these cells were exposed to varying concentrations of Far-C (15, 30, and 60 µM) for 24 h. Subsequently, the media in each well was decanted and the wells were supplemented with 10 µL MTT (5 mg/mL) and the plate was incubated at 37 °C for another 4 h. Thereafter, 100 µL DMSO was further supplemented in each well and the plate was left for 30 min at 37 °C in darkness. Finally, the absorbance of solubilized formazan crystals was recorded at 570 nm (Bio-Rad spectrophotometer, Hercules, CA, USA). Far-C instigated cytotoxicity on HepG2 cells was expressed in terms of cellular viability percentage and was calculated as
$$\mathrm{Cellular}\mathrm{viability}\%=\frac{AbsorbanceofTreatedcells}{AbsrobanceofUntreatedcontrolcells}\times 100.$$
The morphology of HepG2 cells post-exposure with Far-C cells was visualized using microscope. The same procedure as mentioned above was repeated and HepG2 cells were exposed to 15, 30, and 60 µM Far-c and incubated for 24 h. Post incubation, the changes in the morphology of HepG2 cells were visualized under bright light of Floid imaging station (Thermo-Fischer Scientific, Waltham, MA, USA).