The forward primer was (5-GCGGCCGCCAGCTT TCTAGA atggctgggaaaccgaag-3, the underlined was XbaI (Vazyme Biotech Co., Ltd., Nanjing, China) site), and the reverse primer was (5-GAGATGAGTTTTTGT TCTAGA tta ATGATGATGATGATGATG gaaactgaatatttt-3, the underlined was XbaI site, the italic was 6 His tag)
A subcutaneous (under-the-skin) vial format exists, but the dose volume math makes injection awkward at common doses
All animals were aged for at least 12 weeks before being used in their respective experiments
Cayman Chemical, Ann Arbor, MI, USA) according to the manufacturers instructions
Melanotan II was a cyclic form with dramatically higher potency and non-selective activity at all melanocortin receptors