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. 2024 Feb 7;15:1148. doi: 10.1038/s41467-024-45324-w

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a (left) XTT assay (5 days) and (right) clonogenic assay (14 days) of EV/FUT4-OE WM793 cells treated ± 10 μM ARi or cultured in 10% CSS (XTT assay: n = 6 biologically independent samples; clonogenic assay: representative images are shown for 3 independent experiments). b XTT assay of shNT/shFUT4 WM793 cells treated ± 100 nM DHT for 7 days (n = 12 biologically independent samples). c Scratch migration assays of EV/FUT4-OE WM793 cells treated ± 250 μM 2F-peracetyl-fucose (2FF) for 3 days (n = 24 scratches examined over 3 independent experiments). d Scratch migration assay (upper: EV-CTL/ARi, n = 14 scratches; FUT4-OE-CTL/ARi, n = 18 scratches examined over 3 independent experiments. lower: shNT-CTL/DHT, n = 26 scratches; shFUT4-CTL, n = 26 scratches; shFUT4-DHT, n = 25 scratches examined over 3 independent experiments) and e Matrigel invasion assay (upper: EV-CTL, n = 15 fields; EV-ARi, n = 17 fields; FUT4-OE-CTL/ARi, n = 17 fields examined over 2 independent experiments. lower: shNT-CTL, n = 19 fields; shNT-DHT, n = 17 fields; shFUT4-CTL, n = 17 fields; shFUT4-DHT, n = 18 fields examined over 3 independent experiments) of EV/FUT4-OE WM793 cells treated ± 10 μM ARi for 48 h (upper) or shNT/shFUT4 WM793 cells treated ± 100 nM DHT for 48 h (lower). Scale bar = 200 μm. f Scratch migration assay of FUT4/FUT8 double-modified WM793 cells. n = 16 scratches examined over 3 independent experiments. Scale bar = 400 μm. g FITC-gelatin degradation assay of EV/FUT4-OE WM793 cells treated ± 10 μM ARi for 48 h (left; n = 10 fields examined over 3 independent experiments) or shNT/shFUT4 WM793 cells treated ± 100 nM DHT for 48 h (right; shNT-CTL, n = 6 fields; shNT-DHT, n = 9 fields; shFUT4-CTL/DHT, n = 8 fields examined over 3 independent experiments). h 3D spheroid cell invasion assay with EV/FUT4-OE WM793 cells treated ± 10 μM ARi for 7 days (EV-CTL, n = 7; EV-ARi, n = 3; FUT4-OE-CTL, n = 4; FUT4-OE-ARi, n = 4 biologically independent samples). Scale bar = 200μm. i Comparison of FUT4 mRNA levels between primary vs. metastatic melanomas in TCGA_SKCM and GSE8401 datasets. j GSEA illustrating the association of FUT4 expression with Hallmark_Epithelial_Mesenchymal_Transition and Jaeger_Metastasis_Up gene signatures in TCGA_SKCM samples. p-values are calculated by two-sided permutation test. FDR, false discovery rate. NES, normalized enrichment score. For a–i, data are presented as mean values ± SEM and p-values are calculated by two-sided Student’s t-test. Source data are provided as a Source Data file.