V 630 bio uv vis

For the detection of reactive oxygen species (ROS), the assays were performed using Rose Bengal as the synthesizer for ROS. This dye, when irradiated with green light, is able to produce singlet oxygen molecules. As a control for the presence of ROS species 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) was used. 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) is widely used in antioxidants studies as the reporter for oxidative stress environment [12 (link),13 (link)]. In a typical experiment, 90 µL of protein stabilized metal NCs at 40 μM in phosphate-buffered solution were mixed with 10 µL of Rose Bengal at 100 mM. The fluorescence was measured in a quartz cuvette before and after the irradiation with a green lamp during 15 min steps for a total time of 60 min. In parallel, 25 µL of ABTS were mixed with 10 µL of Rose Bengal (100 mM) and 65 µL of phosphate-buffered solution as a positive control of ROS detection. Absorbance was measured using a Jasco spectrophotometer (model V630BIO UV-Vis) before and after irradiation with a green LED lamp during 15 min steps for a total time of 60 min.

Intracellular HK and PDH activity was evaluated with a spectrophotometer (V-630 Bio UV-Vis: JASCO, USA) using Quantification Kit, Cat. No: MAK091-1KT and MAK183-1KT, respectively according to the manufacturer’s instructions (Merck KGaA, Sigma-Aldrich, Darmstadt, Germany). The HK and PDH concentrations were determined with OD values at 450 nm.

After the incubation of MCF-7 and MDA-MB-231 with DDM for 24 h and then exposing them to RT, the measurement of glucose, lactate, and HA metabolism in treated and vehicle-treated MCF-7 and MDA-MB-231 cells was performed. Cell supernatants were harvested and indices were measured using commercial kits (Cat. No: GAGO20 and MAK064 (Sigma-Aldrich) for glucose and lactate, respectively) and HA (Cat. No: 029-001, Corgenix, Inc., Peterborough Cambridgeshire, UK) using a spectrophotometer (V-630 Bio UV-Vis, JASCO, Easton, MD, USA). Glucose, lactate, and HA concentrations were determined at 540 nm, 570 nm, and 450 nm, respectively.