Human melanoma specimens were from the Sheba Medical Center (Helsinki ethical approval Smc-8333-10) and the Wolfson Medical Center (Helsinki ethical approval 0015-16-WOMC). Melanoma margins were determined by pathologists. The distance between the epidermis (stratum corneum) and the dermal adipocytes was measured using the ruler tool in the Aperio ImageScope software (unit: μm) at five random places throughout the tissue section, and these distances were averaged. Melanocytic nest numbers and sizes (in μm2) were calculated by analysis of Melan-A–stained patient samples within a 6-mm range. H&E staining was conducted as described in the previous section. Immunostaining was performed as previously described (14). In brief, slides were incubated, according to the manufacturer’s instructions, with FSP1 (ab27957, Abcam), perilipin1 (ab61682, Abcam), IL-6 (ab6672, Abcam), HMB45 (ab732, Abcam or Dako), MART-1/Melan-A (A103, BioSB), and S100 (ab52642, Abcam) antibodies, followed by incubating with the associated fluorophore-conjugated secondary antibodies: Alexa Fluor 488 (A11055, Invitrogen), Alexa Fluor 594 (8889, Cell Signaling), Alexa Fluor 594 (A21203, Invitrogen), or Alexa Fluor 647 (A31571, Invitrogen). Images were obtained at ×4, ×10, and ×20 magnification using fluorescence microscopy (Nikon).

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