(a) IHC analysis of GCNT2 expression in patient melanomas. GCNT2
expression score on melanoma (n = 64) (range = 0–3: 0 = no staining, 1 =
light staining, 2 = moderate staining, and 3 = dark staining) plotted against
patient mortality outcome. (b) Representative IHC image.
(c) ELISA of Gal-8 was performed on sera from normal healthy
volunteers (n = 5) and patients with MM (n = 13). RT-qPCR analysis of GCNT2 in
GCNT2-OE and -KD (d) A375 and (f) A2058 cells. Flow
cytometry of NGFR on (e) GCNT2 EV and OE A375 cells and on
(g) GCNT2 Scr and KD A2058 cells. In vivo limiting dilution
assay of (h) A375 EV/OE cells and (i) A2058 Src/KD
using cell numbers ranging from 105 to 103. At least four
biological replicates were performed. ***P < 0.001,
**P < 0.01, and *P < 0.05.
EV, empty vector; Gal-8, galectin 8; GCNT2, β1,6
N-acetylglucosaminyltransferase 2; IHC, immunohistochemistry; KD, knockdown; MM,
metastatic melanoma; OE, overexpressed; Scr, scrambled; TIC, tumor-initiating
cell.