Fig. 6

WNT5B controls the phenotype of BLBC by activating canonical and non-canonical WNT signaling. a Western blot showing Wnt5b upon knockdown of WNT5B by lentivirus-mediated shRNA in Hs-578 T, MDA-MB-231, and Bcap-37 cells. b Migration and invasion of sh-control with or without Wnt5b ligand and Wnt5b-KD Bcap-37 cells and MDA-MB-231 cells were measured using transwell chamber assays (scale bar = 100 μm). c Data represent the average cell number from 5 viewing fields (B-37: Bcap-37; M-231: MDA-MB-231; **p < 0.005, ***p < 0.001, ****p < 0.0001). d Active β-Catenin (Non-phospho-ser45 and Non-phospho-Ser33/37/Thr41 and total β-Catenin were analyzed by western blot (Hs-578 T and MDA-MB-231 cells were pretreated with Wnt3a, Wnt5a, or Wnt5b; S: Short exposure; L: Long exposure). e Protein level of phospho-JNK (Thr183/Tyr185), JNK, phospho-Erk1/2 (Thr202/Tyr204), Erk1/2, phospho-STAT3 (Tyr705), and STAT3 were analyzed by western blot (Hs-578 T and MDA-MB-231 cells were pretreated with Wnt3a, Wnt5a, or Wnt5b). f Protein level of canonical and non-canonical markers in WNT5B-KD Hs-578 T and MDA-MB-231 cells was analyzed by western blot. g Protein level of EMT markers in WNT5B-KD Hs-578 T and MDA-MB-231 cells was analyzed by western blot. h Protein level of luminal and basal-like markers in WNT5B-KD Hs-578 T and MDA-MB-231 cells was analyzed by western blot. i Cytoskeleton F-actin proteins were stained with phalloidin and viewed under a confocal microscope (scale bar = 20 μm). j Protein expression of phospho-STAT3 (Tyr705) in representative Her-2 positive, luminal A, luminal B and BLBC tissues by IHC (Red scale bar = 200 μm; Purple scale bar = 40 μm). k The association of phospho-STAT3 (Tyr705) expression between basal-like and non-BLBC tissue subtypes were assessed using Pearson’s χ² test (***p = 0.0003)