Fig. 6: Cells isolated from VIVA1-derived primary or metastatic tumours retain increased invasive properties compared to parental MDA-MB-134VI cells.

Cells were isolated from primary tumour or splenic metastatic sites from animals intraductally injected with VIVA1 cells and expanded in vitro. a Cell growth over time was measured in 2D in vitro culture following viable cell counting with AlamarBlue as described in 'Methods'. All cell lines tested showed similar growth kinetics with the graph representing the mean and standard error of eight technical replicates in each of three biological replicates. b Cell invasion was tested using Matrigel-coated invasion chambers and incubation for 7 days, at which time membranes were isolated, stained and invaded cells counted as described in 'Methods'. Graph is the mean and standard error of duplicate wells in each of three independent biological replicates. VIVA1 (P = 0.051) and primary tumour-derived VIVA1-LIG43 cells retained increased invasive abilities compared to MDA-MB-134VI parental cells. **P value <0.001. c RNA isolated from the various cell lines was subjected to qRT-PCR for highly differentially expressed genes identified in previous RNA-seq experiments. Graphs represent mean and standard error for relative levels of expression of various targets as indicated following normalisation to levels of β-actin as a control from three technical replicates for each of three independent biological replicates. *P value < 0.05; **P value <0.001.