Journal: PLOS ONE

Article Title: Cadherin-11 contributes to the heterogenous and dynamic Wnt-Wnt-β-catenin pathway activation in Ewing sarcoma

doi: 10.1371/journal.pone.0305490

Figure Lengend Snippet: (A) β-catenin was stably knocked down by shRNA (KD#1 and KD#2), and was confirmed by immunoblotting. Control is shRNA targeting GFP (CTRL). (B) Effect of β-catenin shRNA-knockdown significantly decreased colony formation in clonogenic assays. Shown are quantification of colonies per wells, as well as representative wells. (C) In non-adherent soft agar colony formation assays, β-catenin knockdown led to a significant decrease in colony formation. (D) in vivo tail vein injection models demonstrate that β-catenin knockdown leads to less metastatic dissemination to the liver as well as lungs. Quantifications represent mean and standard error of the mean from three or more experiments performed independently. Statistical significance was determined using a Mann-Whitney Test with multiple comparisons. ns, not significant, *p<0.05, **p<0.01, ****p<0.0001.

Article Snippet: For β -catenin knockdown, cell lines were transduced with pLKO.1.-puro lentiviral short hairpin RNAs vectors targeting β- catenin expression (#19761and #19762, Addgene) using a 3 rd generation packaging plasmid, pMDLg/pRRE (#12251, Addgene), pRSV-Rev (#12253, Addgene) and pMD2.G (#12259, Addgene).

Techniques: Stable Transfection, shRNA, Western Blot, In Vivo, Injection, MANN-WHITNEY