M5e microplate reader

MCF-7, MDA-MB-231, Caco-2, hMSC, HFF-1 and HUVEC were treated with ELRs at three different concentrations (0.25, 0.5 and 1 mg/mL) for different times (30, 60, 90 and 120min).
Live and dead staining (LIVE/DEAD Viability/Cytoxicity Assay Kit, Invitrogen) was used according to the manufacturer's instructions. Briefly, a stock solution of the LIVE/DEAD reagents (1 μM calcein AM and 2 μM EthD-1 in 10 mL of DPBS) was prepared, samples (100 µL) were distributed in each well and incubated for 20 minutes in the dark, then the fluorescence intensity emission was measured at 525 and 645 nm after excitation at 485 and 525 nm (SpectraMax M5e Molecular Devices microplate reader). Additionally, photographic images of cultures were taken using a Nikon eclipse Ti-SR (Japan) fluorescence microscope.
Three independent experiments, each in triplicate, were performed.

Caco-2 and MDA-MB-231 cells were incubated with complete medium containing nanoparticles at 0.5 mg/mL for 2 hours. FITC-conjugated annexin V and propidium iodide staining (Annexin V FITC Assay Kit, Cayman Chemical) was used according to the manufacturer's instructions, and the fluorescence intensity emission was measured (SpectraMax M5e Molecular Devices microplate reader) at 535 and 595 nm after excitation at 488 and 560 nm, respectively. Three independent experiments, each in triplicate, were performed.