Beyoclick edu 594 in vitro imaging kit

Cell viability assays were performed as previously described [47 (link)]. Briefly, 5 × 10³ cells/well were seeded in 96-well plates. Twenty-four hours after seeding, cells were transfected with siRNA or treated with the indicated drugs for 48 h. Absorbance was measured at the indicated time points. Cell proliferation was quantified based on the incorporation of 5-ethynyl-2’-deoxyuridine (Edu) into DNA by using a BeyoClick™ EdU-594 In Vitro Imaging Kit (Beyotime, Nantong, China) as previously described [48 (link)]. A laser scanning confocal microscope (CLSM, Carl Zeiss LSM800) was used to determine the proportion of nucleated cells that had incorporated Edu. The assay was performed in triplicate and repeated three times in independent experiments.

The cell viability assay was performed as previously described^{35 (link)}. Briefly, 2 × 10³ cells/well was seeded in 96-well plates (n = 6 per group, n = 3, biological replicates). Twenty-four hours after seeding, the cells were transfected with siRNA, or treated with various concentrations of gefitinib or gefitinib + 1,25(OH)₂D₃ in medium containing 10% FBS for 48 h. Absorbance was measured at indicated time points. Cell proliferation was quantified based on the incorporation of 5-ethynyl-2’-deoxyuridine (Edu) into DNA using a BeyoClick™ EdU-594 In Vitro Imaging Kit (Beyotime, Nantong, China) as previously described^{36 (link)}. A laser scanning confocal microscope (Carl Zeiss, Jena, Germany) was used to determine the proportion of nucleated cells that had incorporated Edu. The assay was performed in triplicate and repeated three times in independent experiments.