Additional file 1: of WNT5B governs the phenotype of basal-like breast cancer by activating WNT signaling

Figure S1. Expression of canonical breast cancer subtype-markers in breast cancer. a Expression of selected canonical breast cancer subtype-marker mRNAs with Her-2 positive-specific, luminal-specific, or TNBC-specific based on UALCAN (Red p: the former > the latter; Green p: the former < the latter). b Common breast cancer subtype-marker expression in representative Her-2 positive (Her-2+), luminal A (ER/PR+, Her-2-, EGFR-, Ki-67% < 14%), luminal B (ER/PR+, Her-2-, EGFR-, Ki-67% ≥ 15%), and BLBC (ER-, Her-2-, EGFR+, CK5/6+) tissues by IHC (Red scale bar = 200 μm; Purple scale bar = 40 μm). Figure S2. Identification of BLBC-specific Wnt ligands based on online platforms. a mRNA expression of selected canonical WNT signaling targets with TNBC or non-luminal-specific based on UALCAN (Red p: the former > the latter; Green p: the former < the latter). b mRNA expression of WNT3A, WNT3, WNT5A, WNT5B, and WNT11 in five different breast cancer subtypes based on bc-GenExMiner v4.1 according to the Sørlie’s subtypes (****p < 0.0001; Red star: the former > the latter; Green Star: the former < the latter). c mRNA expression of WNT3A, WNT3, WNT5A, WNT5B, and WNT11 in normal breast, Luminal, Her-2 positive, and TNBC subtypes based on UALCAN (Red p: the former > the latter; Green p: the former < the latter). Prognostic value of WNT3 d and WNT11 e mRNA levels in human breast cancer, data obtained from the KM-plotter. f WNT3A, WNT5A, and WNT5B mRNA expression levels across various normal tissues based on GTEx which were deposited in the HPA. Figure S3. Analysis of canonical WNT signaling constitutive components in breast cancer. a Expression levels of canonical WNT signaling constitutive components in two normal breast cell lines, eight luminal, and seven BLBC cell lines by western blot (S: Short exposure; L: Long exposure). b Expression of selected canonical WNT signaling constitutive component mRNAs with luminal or basal-like specific based on bc-GenExMiner v4.1 according to the Sørlie’s subtypes (*p < 0.05; ****p < 0.0001; Red star: the former > the latter; Green Star: the former < the latter). c Expression of selected canonical WNT signaling constitutive component mRNAs with TNBC or non-TNBC-specific based on UALCAN (Red p: the former > the latter; Green p: the former < the latter). d Expression of β-catenin and active β-catenin (np-Ser45, np-Ser33/37/Thr41) in representative Her-2 positive, luminal A, luminal B and BLBC tissues by IHC (Red scale bar = 200 μm; Purple scale bar = 40 μm). e Expression of β-catenin in two luminal and two BLBC lines by immunofluorescence (confocal microscopy, scale bar = 20 μm). Figure S4. Identifying inhibitors targeting Wnt5b and CK1α based on online platforms. a PPI pattern of WNT3A, WNT5A, and WNT5B based on STRING. b CSNK1A1 is positively correlated with luminal markers. c CSNK1A1 is inversely correlated with basal-like marker KRT5. d CSNK1A1 is positively correlated with EMT-attenuated markers CDH1 and TJP1. e CSNK1A1 is inversely correlated with EMT acquired markers VIM and SNAI1. f CSNK1A1 is inversely correlated with WNT5B. g CSNK1A1 is positively correlated with CCND1. (b-g) were based on GEPIA (breast invasive carcinoma; n = 1085). h Ultrasound images of BLBC cells bearing mice under B-type ultrasonic image system at day 6, 12, and 18 for Bcap-37 following treatment with LGK-974 (5 mg/kg) or pyrvinium pamoate (1 mg/kg). i Percentage of nuclei positive for Ki67 after 14 days of LGK-974 or pyrvinium pamoate treatment. Graphs represent mean ± SEM (n = 5 per treatment group; ****p < 0.0001, n.s.: no significant). Figure S5. Proposed model of WNT5B governing the phenotype of BLBC by activating canonical and non-canonical WNT signaling. (PDF 14908 kb)