10 μl cck8 solution

Cisplatin-resistant HCC cells (HCC-LM3/DDP and Huh-7/DDP) and HCC cells (HCC-LM3 and Huh-7) were in the logarithmic growth stage and digested with trypsin. Cell number was calculated using Countess™ Cell Counting Chamber Slides (Catalog number: C10228, Invitrogen™). With the cell density adjusted to 2 × 10⁴/mL, 100 μL medium was added to 96-well plates. Cisplatin at different concentrations was administered to treat the cells for 24 h, and the 96-well plates were further cultured in an incubator. A 10 μL CCK8 solution (Beyotime Biotechnology, Shanghai, China) was added to each well after 24, 48, or 96 h to incubate the plates for another 1 h in the incubator. As the culture ended, a microplate reader in which the 96-well plates were placed was exploited to check the absorbance (OD value) of each well at 450 nm, after which the determination was taken at the 24th, 48th, 72nd, and 96th hours.

The CCK-8 assay was used to measure the viability of LLC cell treated with abPTX or NP-abPTX. Briefly, LLC cells in the exponential growth phase (3000 cells/well) were cultured in 96-well plates (Corning, Corning, NY) for 24 h, and were then treated with abPTX or NP-abPTX for 48 and 72 h at 37 °C, respectively. Nontreated group and abPTX-treated group were considered as the control group and positive group, respectively. Each group was repeated four times. For detecting the optical density (OD), 10 μL CCK-8 solution (C0038, Beyotime, Shanghai, China) was subsequently added to each group and was incubated with LLC cells for 2 h at 37 °C. The OD value was measured by Cytation 3 Cell Imaging Multi-Mode Reader (BioTek, Winooski, VT) at a wavelength of 450 nm. Relative cell viability was calculated using the following formula: relative cell viability=(mean OD450 of experimental groups/mean OD450 of control groups × 100%).