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High Fat Diet + 2% freeze dried unroasted caffeinated coffee ( HFGC )

Transcriptomic Analysis . 9 weeks. n=4. CON Diet ( D12450B , Research Diets, ND). n=4. High Fat Diet (D12492 , 60% kcal fat, HF). n=5. High Fat Diet + 2% freeze dried caffeinated coffee (HFCC). n=5. High Fat Diet + 2% freeze dried decaffeinated coffee ( HFDC ). n=5.

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High Fat Diet + 2% freeze dried unroasted caffeinated coffee ( HFGC )

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  1. Transcriptomic Analysis 9 weeks n=4 CON Diet(D12450B, Research Diets, ND) n=4 High Fat Diet (D12492, 60% kcal fat, HF) n=5 High Fat Diet + 2% freeze dried caffeinated coffee (HFCC) n=5 High Fat Diet + 2% freeze dried decaffeinated coffee (HFDC) n=5 High Fat Diet + 2% freeze dried unroasted caffeinated coffee (HFGC) Dissection Acclimatization Plasma • Measurement of plasma triglycerides and total cholesterol levels • Extracted mRNA from each respective pool was pooled and subjected to microarray analysis (Mouse Genome 430 2.0 Array) • Real-time reverse transcription PCR on individual samples to detect gene expression levels • Extracted protein from individual samples to measure Atf3 and Socs3 protein levels Skeletal Muscle

  2. Insulin Signalling 8 n=9 CON Diet(D12450B, Research Diets, ND) n=9 High Fat Diet (D12492, 60% kcal fat, HF) n=9 High Fat Diet + 2% freeze dried caffeinated coffee (HFCC) n=9 High Fat Diet + 2% freeze dried decaffeinated coffee (HFDC) n=9 High Fat Diet + 2% freeze dried unroasted caffeinated coffee (HFGC) Dissection 8th week: Insulin tolerance test Acclimatization In vivo stimulation: saline (n=4), insulin (n=5) Excised skeletal muscle and extracted protein Measurement of protein levels of p85/IRS-1; tyrosine phosphorylated IRS-1 and ser473 phosphorylated Akt/PKB

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