Synergy h1 automatic microplate reader

HAECs (5 × 10³ cells/well) were seeded in 96-well plates. After adherence, the cells were incubated in ECM with or without HG/PA and GSC extracts. After 48 h of incubation, 10 μL of CCK-8 (Dojindo, Japan) solution and 90 μL of ECM were added to each well, and the cells were further incubated in the CO₂ incubator at 37°C for 2 or 3 h. The absorbance was measured at 450 nm using a Synergy H1 Automatic Microplate Reader (BioTek, USA).

The cell viability of GC, MSN-NH₂, MSN-NH₂/GC, 5-FU, 5-FU@MSN-NH₂, 5-FU@MSN-NH₂/GC to SW620 cells was assessed by the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cells were seeded in a 96-well plate (2 × 10⁴cell/well) and incubated for 24 h at 37°C with 5% CO₂. Afterwards, the culture medium was replaced with a fresh medium containing the GC, MSN-NH₂, MSN-NH₂/GC, 5-FU, 5-FU@MSN-NH₂, 5-FU@MSN-NH₂/GC at different concentrations and incubated for 48 h, with cells treated with medium only as a control. After being cultured for 48 h, the cells were treated with 20 µl MTT (5 mg ml⁻¹ in PBS) for 4 h at 37°C. Then, the medium was removed and 150 µl of dimethyl sulfoxide was added to each well to dissolve the purple formazan. The optical density (OD) of each well was measured at a wavelength of 490 nm by Synergy H1 automatic microplate reader (BioTek, Winooski, VT, USA). The cell viability was calculated.

Mice were administered with 100 μl of 100 mg/ml 70-kDa FITC-dextran (Sigma-Aldrich) in sterile water, and blood was collected 4 h before sacrifice (Wang et al., 2020b (link)). Serum was obtained by centrifugation (3,000 rpm, 10 min, 4°C). The concentration of FITC in serum was determined by SynergyH1 automatic microplate reader (Biotek) with excitation wavelength of 485 nm and emission wavelength of 528 nm (Pu et al., 2021 (link)).