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Guo D, Reinitz F, Youssef M, et al

Melanocytes were maintained in full 2:1 melanocyte growth media (2 parts minimal essential medium (MEM) supplemented with 1% foetal bovine serum (FBS), 1% nonessential amino acids, 100 g/ml Primocin, 2 mM Glutamax, 5 ng/ml basic fibroblast growth factor, and 5 ng/ml endothelin-1 (Sigma, Dorset, UK) mixed with 1 part keratinocyte serum-free medium (K-SFM) supplemented with 25 g/ml bovine pituitary extract (BPE), 0.2 ng/ml rEGF, 100 g/ml Primocin (antibiotic), and 2 mM Glutamax in humidified incubators at 37 C with 5% CO 2 unless stated otherwise

Amplification conditions were optimized for each primer pair, and melt-curve analysis was performed at the end of the reactions to confirm amplification specificity

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These findings underscore oral microbes as upstream regulators of the gutbrain axis and provide a microbialneural perspective on the pathological link between periodontitis and depression (Figure 2)