Journal: Scientific Reports
Article Title: Canagliflozin inhibits interleukin-1β-stimulated cytokine and chemokine secretion in vascular endothelial cells by AMP-activated protein kinase-dependent and -independent mechanisms
doi: 10.1038/s41598-018-23420-4
Figure Lengend Snippet: Canagliflozin inhibits 2-deoxyglucose uptake in HUVECs without influencing cell proliferation. ( a ) HUVECs were preincubated in the presence or absence of canagliflozin (10 μmol/l) for the times indicated or dapagliflozin (10 μmol/l, 30 min) and 2-deoxyglucose uptake assessed. Data shown represents the mean 2-deoxyglucose uptake relative to vehicle from three independent experiments. **p < 0.01, ***p < 0.001 relative to vehicle (one-way ANOVA). ( b ) Cell lysates from HUVECs (2 independent lysates), HAECs (4 independent lysates) and HEK-293 cells and mouse kidney membranes were resolved by SDS-PAGE and immunoblotted with anti-SGLT2 antibodies. The immunoblot shown has been cropped, with the full-length immunoblot shown in Supplementary Figure . ( c , d ) HUVECs were incubated in the presence or absence of canagliflozin, dapagliflozin or empagliflozin and ( c ) proliferation assessed in 2.5% (v/v) or 5% (v/v) MV2 Supplement mix (serum) by BrdU incorporation or ( d ) viability assessed by MTS assay. Data shown represents the mean proliferation or viability relative to vehicle (in 5% MV2 supplement mix) from three independent experiments in each cae. ## p < 0.01 relative to vehicle (two-tailed t-test).
Article Snippet: HUVECs (2 × 10 4 cells/well) were allowed to attach to 96-well plates for 4 h in MV2 medium containing 2.5% or 5% (v/v) MV2 Supplement Mix (Promocell C-39226) containing serum and growth factors and subsequently incubated in the same medium with DMSO (vehicle) or SGLT2 inhibitors for 12 h. BrdU reagent was then added and cells further incubated for 24 h. BrdU incorporation was assessed by measuring A 485/520 on a BMG Labtech FLUOstar Optima spectrophotometer microplate reader.
Techniques: SDS Page, Western Blot, Incubation, BrdU Incorporation Assay, MTS Assay, Two Tailed Test
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