Fig. 4
Knockdown of galectin-3 (Gal3) reduces canonical Wnt and AKT signaling in CSC-like spheres. a Canonical Wnt signaling activity was evaluated by SuperTOP/FOP (STF) assays and normalized to Renilla luciferase (RLU = relative light units). Baseline Wnt activity was reduced in Gal3negative GI-LM2G spheres (grey column) compared to Gal3positive GI-LM2C spheres (white column). b Flow cytometric analysis of the Wnt target gene LGR5 in GI-LM2C (red) and GI-LM2G (green) spheres. Canonical Wnt activity in (a) correlated with LGR5 expression. c TGFβ signaling was evaluated by SBE4-luc assays measured as normalized firefly to Renilla luciferase activity (RLU = relative light units). GI-LM2C (red column) and GI-LM2G (green column) revealed no significant difference in TGFβ activity. d Western blot analysis shows typical results for EMT markers (E-cadherin, vimentin), Gal3 as well as phospho-AKT/total-AKT in GI-LM2C and GI-LM2G spheres. P-AKT was absent in GI-LM2G. Experiments were carried out in triplicate (n = 3)