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AFM-based hydrophobicity mapping further showed that incubating cells with Cu decreased the surface hydrophobicity with an increase of incubation time.
Red blood cells were removed by incubating cells in 1× RBC lysis buffer (BioLegend) for 5 min at room temperature.
After incubating cells, the MTT treatment solution was carefully and completely aspirated from wells, with DMSO then added (180 µL) to extract cellular Formazan.
The protease inhibitors lethal dose was determined by incubating cells with different concentration of the protease inhibitor and evaluating their effect on cell viability.
The cells were gently removed as described12 or alternatively by incubating cells with 20 mM NH4OH, 5 mM EDTA at RT with gentle swirling of the plate so that the intact ECM remained on the coverslips.
Appropriate controls were performed by incubating cells with the corresponding concentrations of glycol-chitosan, but without BTNPs.
Figure 7a, b illustrated that incubating cells with NPs up to 48 h, maximum (61%) G1/S blocking occurred as compared with native etoposide (52%).
Unspecific binding sites were subsequently blocked by incubating cells with 5% goat serum.
For ELISA, cell culture supernatant was harvested after incubating cells with peptides for 24 h.
Starvation of serum, glucose and amino acids was carried out by incubating cells in DPBS.
Serum starvation was carried out by incubating cells in their corresponding medium lacking fetal bovine serum.
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