Brdu colorimetric assay kit

HUVEC cells (7 × 10³/well), were cultured into 96-well plates for 24 h before the addition of HT at the indicated concentrations and cultured for additional 24–48 h at 37 °C. Cell proliferation was evaluated by measuring BrdU incorporation into DNA (BrdU colorimetric assay kit; Roche Applied Science, South San Francisco, CA, USA), by an ELISA plate reader (Thermo Scientific, Waltham, MA, USA) at 450 nm, as described in detail elsewhere [65 (link)]. In all experiments, performed in triplicate, the relative cell growth was expressed as percentage in comparison with untreated 24 h control cells (100%).

HaCaT cells (6 × 10³/well) were cultured for 24 h into 96-well plates before the addition of HT at the indicated concentrations and were cultured for an additional 24–48 h at 37 °C. BrdU incorporation into DNA (BrdU colorimetric assay kit; Roche Applied Science, South San Francisco, CA, USA) was used to measure cell proliferation, determined by an ELISA (enzyme-linked immunosorbent assay) plate reader (ThermoScientific, Waltham, MA, USA) at 450 nm as described in detail elsewhere [68 (link)]. All experiments were performed in triplicate, and the relative cell growth was expressed as a percentage in comparison with the untreated control cells.

U87-MG cell proliferation was evaluated by measuring BrdU incorporation into DNA (BrdU colorimetric assay kit; Roche Applied Science, South San Francisco, CA, USA) following treatment with temozolomide (100 μM) combined with increasing doses of LAV-BPIFB4 (0–144 ng/mL) for 96 h or following treatment with LAV-BPIFB4 at 18 ng/mL without temozolomide. Newly synthesized BrdU-DNA was determined on an ELISA plate reader (Thermo Scientific, Waltham, MA, USA) at 450 nm. All experiments were performed in triplicate, and the relative cell growth was expressed as a percentage of the untreated control cells (set to 100%) to allow for an unwanted source of variation.

HaCaT cells (6 × 10³/well) were cultured for 24 h into 96-well plates before the addition of the G. lucidum ethanol extract or ganoderic acid-A at the indicated concentrations and were cultured for an additional 24–48 h at 37 °C. Cell proliferation was evaluated by measuring BrdU incorporation into DNA (BrdU colorimetric assay kit; Roche Applied Science, South San Francisco, CA, USA) and was determined by an ELISA plate reader (ThermoScientific, Waltham, MA, USA) at 450 nm as described in detail elsewhere [43 (link)]. All experiments were performed in triplicate, and the relative cell growth was expressed as percentage in comparison with the untreated control cells (100%).