Detecting mRNA expression in melanoma cells

Antisense single-stranded DNA probe (Supplementary Table S3) was synthesized and end-labeled with digoxigenin (DIG) (Roche). ISH or FISH was performed in formalin-fixed paraffin-embedded melanoma sections or slides covered with cultured melanoma cells. The pre-hybridization, hybridization, anti-DIG-HRP IgG fraction monoclonal (Jackson, 200-032-156) incubation (1:200), and stained with DAB (Servicebio, G1211) was performed as described in previous studies [38 (link), 39 (link)]. Stained ISH or FISH sections were imaged with an Olympus FV1000 Confocal Laser Scanning Microscope or a ZEISS Axio Vert.A1 microscope and at least 10 representative images were collected for statistical analysis. The ISH or FISH staining was performed “blind” with respect to the different treatments.