Xtt cell proliferation assay kit

To check the cell proliferation, XTT cell proliferation assay kit (Sigma®, St Louis, Missouri, USA) was used. This kit measures the cell proliferation based upon the reduction of the tetrazolium salt, 2,3-Bis (2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide [19] (link). After Concanavalin A (ConA) stimulation for 48 h, XTT reagent was added in each well and cells were further incubated for 20 h at 37 °C in CO₂ incubator. After incubation, the plate was gently shaken on the rocker to evenly distribute the dye in the wells. Using ELISA plate reader, absorbance was measured at 450 nm considering 650 nm wavelength as reference.

Silver nitrate (Ag(NO₃)), sodium boron hydride (NaBH₄), trisodium citrate, dimethylsulfoxide (DMSO), and XTT cell proliferation assay kit were purchased from Sigma (St. Louis, USA). Dulbecco's Modified Eagle's Medium (DMEM), penicillin-streptomycin, trypsin, and fetal bovine serum (FBS) were purchased from Gibco (USA). 20x phosphate-buffered saline (PBS) was purchased from Thermo Fisher (USA), cation-adjusted Mueller-Hinton broth (CAMHB) and 2,3,5-triphenyltetrazolium chloride (TTC) were purchased from Merck (Germany), and annexin V/PI assay kit BD (USA) and paclitaxel were purchased from Sandoz (Germany).