-values 0.1). Genetic instruments for type 2 diabetes were obtained from the latest GWAS on incident type 2 diabetes
. Horizontal pleiotropy was estimated based on MR-Egger intercept. Cochran Q-statistic was used to estimate heterogeneity of instruments.HEK-Blue IL-18 cells passages 1-16 were cultured in 4.5 g/L glucose Dulbecco’s Modified Eagle’s Medium supplemented with 10% foetal bovine serum , 2 mM L-Glutamine, 50 U/ml penicillin, 50 μg/ml streptomycin and 100 μg/ml Normocin . HEK-Blue IL-18 cells were designed to detect bioactive IL-18 by monitoring the activation of the NF-κB and AP-1 pathways.
Murine C3H10T1/2 cells were seeded at 100,000 cells/well in 12 well dishes in DMEM/F12 media supplemented with 10% FBS, 1% P/S. After 48 h, when cells reached 100% confluency, media was replaced with differentiation media DMEM/F12 containing 2 µg/mL Insulin, 0.5 mM IBMX, 2 ug/mL Dexamethasone, and 5 µM Rosiglitazone for 48 hrs. Media was then replaced with maintenance media containing DMEM/F12 supplemented with 2 µg/mL insulin for 5 days.
HepG2 Cells were cultivated in DMEM media with 1 g/L of glucose, supplemented with 10% SVF and maintained at 37 °C and 5% CO2. Cells were plated in DMEM media with 10% SVF.Primary hepatocytes and C3H10T1/2-derived adipocytes were treated with NogoR recombinant protein for 3 or 6 h respectively prior to 100 nM insulin stimulation for 15 min. HepG2 cells were treated with 0, 1 nM, 10 nM, or 100 nM of NogoR or CRELD1 recombinant proteins for 3 h and then stimulated with insulin for 15 min.