Flow cytometry analysis of rhGal-8 binding on (a) A375
GCNT2 EV/OE and (b) A2058 GCNT2 Scr/KD cells. GST-tagged Gal-8
affinity chromatography followed by immunoblotting for CD44 on parental
(c) A375 and (e) A2058 cells, PNGase-treated and
control parental (d) A375 and (f) A2058 cells, and
(g) A375 GCNT2 EV/OE and (h) A2058 GCNT2 Scr/KD
cells. Immunoblot analysis of Akt activation after Gal-8 treatment on
(i) A2058 GCNT2 Scr/KD cells. At least four biological
replicates were performed. ****P < 0.0001,
***P < 0.001, **P < 0.01,
and *P < 0.05. Akt, protein kinase B; EV, empty vector;
Gal-8, galectin 8; GCNT2, β1,6 N-acetylglucosaminyltransferase 2; GST,
glutathione S-transferase; KD, knockdown; MM, metastatic melanoma; OE,
overexpressed; poly-LacNAc, poly-N-acetyllactosamine; rhGal-8, recombinant human
galectin 8; Scr, scrambled.