Nature Medicine: doi: /nm.3891
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1 Supplementary Figure 1. Subjective responses. Thermal sensation, thermal comfort and self-reported shivering, determined at several time points (from t = min until t = 36 min) after entering the cold room, on day 1 compared to day 3 of the cold acclimation period (P values for iaucs:.36,.69 and.17, respectively). n = 8; comparisons between day 3 and day 1 were analyzed using Wilcoxon signed-rank tests; *P <.5, day 3 vs. day 1 for specific time point; error bars, mean ± s.e.m. Nature Medicine: doi:1.138/nm.3891
2 IMCL area fraction (%) Sarcolipin (AU) Serca2 (AU) OXPHOS proteins (AU) PGC-1a (AU) a b c d State 2 State 3 - fat oxidation State 3 respiration - complex I+II 8 State U M MO MO MOG MOGS MOGS e f g State 4o respiration II III IV V h i j Total Type 1 Type 2 Sarcolipin PGC-1α Serca Supplementary Figure 2. Skeletal muscle mitochondrial respiration, intramyocellular lipid content, and OXPHOS, PGC-1α, Serca2 and sarcolipin protein content. (a-e) Respiration measurements in permeabilized muscle fibers. M, malate; O, octanoyl carnitine; P, pyruvate; G, glutamate; S, succinate. (a) Respiration upon substrates only (state 2). (b,c) ADP-stimulated respiration fuelled by mitochondrial complex I and complex II-linked substrates. (d) Maximally uncoupled respiration upon addition of the chemical uncoupler FCCP (state U, maximal electron transport chain capacity). (e) Oxygen consumption not linked to ATP synthesis upon addition of the ATP-ase inhibitor oligomycin (state 4o, mitochondrial leak respiration). (f) Expression of oxidative phosphorylation protein (OXPHOS) complexes II through V. (g) PGC-1α protein content, including Nature Medicine: doi:1.138/nm.3891
3 representative Western blots of two individuals with T2D before and after cold acclimation. (h) Total intramyocellular lipid (IMCL) content and IMCL content in type 1 and type 2 muscle fibers. (i,j) Sarcolipin (i) and Serca2 (j) protein content, including representative Western blots of two individuals with T2D before and after cold acclimation. White bars, before cold acclimation; black bars, after cold acclimation. n = 7; comparisons between data before and after cold acclimation were analyzed using Wilcoxon signed-rank tests; error bars, mean ± s.e.m. Nature Medicine: doi:1.138/nm.3891
4 Supplementary Table 1. Subject characteristics. Patient Age BMI Diabetes duration (years) Medication HbA1c (mmol/mol) Metformin (5mg 2x/day), 48.6 Glimepiride (2mg, 1x/day) Metformin (1mg, 2x/day) Metformin (1mg, 2x/day) Metformin (1mg, 2x/day), 62.8 Glimepiride (5mg, 1x/day) Metformin (5mg, 2x/day) Metformin (85 mg, 3x/day), 6.7 Glimepiride (6mg, 1x/day) Metformin (85mg, 2x/day), 54.1 Vildagliptin (5mg, 2x/day) Metformin (85mg, 3x/day), Gliclazide (8mg, 2x/day) 54.1 Nature Medicine: doi:1.138/nm.3891
5 Supplementary Table 2. Body weight and fasting blood parameters before and after cold acclimation. Before After Body weight (kg) 92. ± ± 1.8 Glucose (mmol/l) 7.9 ± ± 3.2 Insulin (pmol/l) ± ± 45.7 NEFA (μmol/l) 644 ± ± 158 Fructosamine (μmol/l) 248 ± ± 25 TSH (mu/l) 2.9 ± ± 1.2 A Free T4 (pmol/l) 12.9 ± ± 1.8 Noradrenaline (pg/ml) 43 ± ± 146 Phosphate (mmol/l) 1.3 ±.1.98 ±.13 Urea (mmol/l) 5.1 ± ± 1.2 Creatinine (μmol/l) 77.3 ± ± 9.4 γgt (U/l) 34.6 ± ± 14.1 ASAT (U/l) 39.1 ± ± 8.7 ALAT (U/l) 4.5 ± ± 26.7 Billirubine (μmol/l) 11.9 ± ± 6.3 Total cholesterol (mmol/l) 3.4 ± ±.9 HDL cholesterol (mmol/l) 1.1 ± ±.2 LDL cholesterol (mmol/l) 1.6 ± ±.9 Triglycerides (mmol/l) 1.67 ± ±.66 Total protein (g/l) 68.4 ± ± 2.7 Albumin (g/l) 37.2 ± ± 2.3 A P <.5, before vs. after cold acclimation. Values are expressed as mean ± s.d. Nature Medicine: doi:1.138/nm.3891
6 Supplementary Table 3. Blood pressure, body temperatures and skin perfusion during thermoneutral and mild cold conditions, before and after cold acclimation. Before After Variable Thermoneutral Mild cold Delta Thermoneutral Mild cold Delta Systolic blood pressure 125. ± ± 13.9 B 24.1 ± ± ± 2.9 B 27.1 ± 11. (mm Hg) Diastolic blood pressure 74.4 ± ± 9.5 B 1.8 ± ± ± 7.4 B 12.8 ± 3.4 (mm Hg) Mean arterial pressure 91.3 ± ± 9.7 B 15.3 ± ± ± 11.3 B 17.5 ± 4.1 (mm Hg) Heart rate (beats/min) 67.1 ± ± 6.9 B 8.8 ± ± ± 5.6 B 7.3 ± 5.3 Suit temperature ( C) 29.9 ± ± ± ± ± ± 1. Core temperature ( C) 36.9 ± ±.4.1 ± ± ±.3.1 ±.3 Mean skin temperature ( C) 35.2 ± ± ± ±.3 A 28.8 ± ± 1.2 Skin perfusion hand ± 7.1 B 9.8 ± ± 23.4 B 85.4 ± 23.4 relative % Skin perfusion underarm relative % ± 12.1 B 66.5 ± ± 24.6 B 66. ± 24.6 A P <.5, before vs. after cold acclimation; B P <.5, thermoneutral vs. mild cold. Values are expressed as mean ± s.d. Nature Medicine: doi:1.138/nm.3891
7 Supplementary Table 4. Substrate kinetics during hyperinsulinemic euglycemic clamp. Before After Rd glucose (μmol kg -1 min -1 ) Delta (Rd clamp Rd basal) 8.1 ± ± ± ± ± 6.9 A 11.6 ± 7.1 A EGP (μmol kg -1 min -1 ) Delta (EGP clamp EGP basal) % Delta 7.2 ± ± ± ± ±1.6.9 ± ± ± 17.5 Carbohydrate oxidation (μmol kg -1 min -1 ) Delta (CHO ox clamp CHO ox basal) 6.7 ± ± ± ± ± ± 2. NOGD (μmol kg -1 min -1 ) Delta (NOGD clamp NOGD basal) 1.4 ± ± ± ± ± ± 6.5 Lipid oxidation (μmol kg -1 min -1 ) Delta (lipid ox clamp lipid ox basal) 1.2 ±.2.8 ±.4.3 ± ±.2.6 ±.2.5 ±.2 A P <.5, before vs. after cold acclimation. Values are expressed as mean ± s.d. Rd, rate of disappearance; EGP, endogenous glucose production; NOGD, nonoxidative glucose disposal. Nature Medicine: doi:1.138/nm.3891
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