Fig. 5

Identification and quantification of SLAMF9⁺ cells in benign and malignant melanocytic lesions. a Western blot analysis of three independent transgenic SLAMF9⁺ U937clones and EV U937 clones was performed to validate specificity of our self-generated polyclonal anti-hsSLAMF9 antibody. The respective guinea pig IgG was used as isotype control and GAPDH was used as a loading control. b Human melanoma specimens were immunohistochemically stained with the self-generated polyclonal anti-hsSLAMF9 antibody and guinea pig IgG isotype control. Representative pictures of pT1 and pT4 melanoma specimens are shown, scale bar = 100 µm. c In situ hybridization was performed to visualize Slamf9 mRNA in formalin-fixed paraffin-embedded melanomas. One representative picture of a pT4 melanoma specimen is shown, scale bar = 100 µm. d Immunofluorescent double staining of SLAMF9 (green) and CD68 (red) as well as the respective guinea pig IgG and mouse IgG isotype controls of a human pT4 melanoma was performed, scale bar = 100 µm. E Epidermis, I inflammatory infiltrate, T Tumor. e Quantification of SLAMF9⁺ cells in tissue microarrays with benign and malignant melanocytic lesions. Two core sections of each specimen were evaluated after immunohistochemical staining of SLAMF9. The percentage of samples containing SLAMF9⁺ TAM only or SLAMF9⁺ TAM plus SLAMF9⁺ tumor cells was histopathologically assessed