Fig. 5

Increased spheroid formation by CPEB2-KO and CPEB2-KD cells. a Representative images of spheroids (Scale = 60 μm) and (b) spheroid forming efficiency (SFE) of WT and CPEB2KO MCF10A cells grown on ultra-low attachment plates for 4 days. SFE is computed as the number of spheroids (> 60 μm) divided by total number of cells plated. CPEB2KO cells showed 5.12-fold increase in SFE (p < 0.001). c Dot plot of spheroid size (Mann-Whitney Test for statistical significance) showing increased average diameter (WT = 70.92 μm, CPEB2KO = 91.42 μm; p < 0.0005), indicating enhanced spheroid growth rate. d Images of spheroids and (e) SFE in Mock and CBEB2-KD MCF7 cells, showing an increase in CPEB2-KD cells. h Representative Western blot and (g) quantification (Mean of 3 ± SEM) of β- Catenin protein expression. f qRT-PCR (Mean of 3 ± SEM) for downstream genes of β-Catenin pathway. β-Catenin was increased 1.29 fold (p = 0.048) in CPEB2KO cells, with increases in downstream target genes CCND1 (3.49 fold, p = 0.039), and AXIN1 (1.298 fold, p = 0.034). No significant change was observed in AXIN2 (1.73 fold change, p = 0.10) or Myc (0.76 fold change, p = 0.051)