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The primers and probes used for Mc4r and the internal control Ngf were as follows (53): -Mc4r Forward: atttgcagcctgctttcca, Reverse: tggagcgcgtaaaagatagtga FAM-MGB Probe: tgcggtggacaggta, -Ngf, Forward: tgcatagcgtaatgtccatgttg, Reverse: tctccttctgggacattgctatc, VIC-TAMRA Probe: acggttctgcctgtacgccgatca (all primers and probes were synthesized by Thermo Fisher Scientific Inc.)