Next, we validated the highest significantly down- (Myh2, Mybh, Sypl2, Xirp1, Mybpc1) and up- (Fcgbp, Areg) regulated genes, including the melanoma-specific genes (Dmnt1 and Nrp2) identified by RNA sequencing following treatment with SAM+anti-PD-1 by RT-qPCR. This evidence concerns the gene AREG and melanoma.