Journal: bioRxiv

Article Title: Cooperation between MYC and β-catenin in liver tumorigenesis requires Yap/Taz

doi: 10.1101/819631

Figure Lengend Snippet: MYC and β-cat Ex3 activation cooperate in transformation in liver and fibroblasts. (A) Kaplan Meyer disease-free survival curves for mice of the indicated genotypes (all in the presence of the Alb-CreER T2 transgene). The number of mice (n) and the median survival are indicated. p-values were calculated with the log-rank test. (B) Western blot analysis of β-catenin and MYC protein expression in wild-type (wt) liver and representative R26-lslMYC and β-cat Ex3 ;R26-lslMYC liver tumor samples. Vinculin was used as loading control. Each tumor is identified by its unique reference number. (C) Hematoxylin and Eosin staining of representative liver sections from the indicated genotypes. Bars: 400 µm (H&E 5x) or 100 µm (H&E 5x). (D) Left: representative pictures of the colony forming assay of 3T9 MycER;S33Y cells plated in 50% (v/v) methylcellulose and cultured for 7-10 days in the presence of 1 μM Doxycycline and/or 400 nM 4-OHT, as indicated. This experiment was performed four times, each with technical triplicates. Bar: 1000 µm. Right: quantification of the total number of colonies with a diameter >10 μM in each condition. Bar plot represent the average and standard deviation for the 4 biological replicates. (E) Representative pictures of CD1-nude mice 6 weeks after injection of 3×10 6 3T9 MycER;S33Y cells and fed with Tamoxifen (Tam) and/or Doxycyline (Doxy), as indicated. The fraction of tumor-bearing mice in each experimental group is indicated below each photograph.

Article Snippet: After addition of 6X Laemmli buffer (375 mM Tris-HCl, 9% SDS, 50% glycerol, 9% beta-mercatoethanol and 0.03% bromophenol blue), lysates were boiled (5 minute at 100°C) and then electrophoresed on pre-cast 4-15% gradient Polyacrilamide gels (Bio-Rad, #5678084), transferred onto methylcellulose membranes (Bio-Rad, #1704271) and protein expression detected with the indicated primary antibodies (Supplementary Table 5).

Techniques: Activation Assay, Transformation Assay, Western Blot, Expressing, Control, Staining, Cell Culture, Standard Deviation, Injection