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The cells were seeded in a 6-well plate (Corning) and transfected with the constructs of rat wild-type Tshr ( Tshr WT -GFP ) and truncated Tshr ( Tshr Df -GFP ) in pAcGFP1-N1 (Clontech, 632469) backbone (The cells transfected with GFP expression vector pAAV-GFP was used as negative control)

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88 As most properly formulated commercial dog foods are fortified to include the recommended levels of vitamins and minerals for healthy dogs at different life stages, supplementation is not necessary for most pets, and in some cases, can be harmful