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. Author manuscript; available in PMC: 2023 Apr 24.
Published in final edited form as: J Invest Dermatol. 2022 Sep 27;143(3):456–469.e8. doi: 10.1016/j.jid.2022.07.033

Figure 5. Gal-8 bound i-linear poly-LacNAcs preferentially on MM cells, and the major Gal-8 ligand on MM cells was identified as CD44.

Figure 5.

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.