Fig. 7. Knockdown of RB1 phenocopies miR-181a mediated transformation.

a Graphs showing RB1 mRNA and protein expression in the FT237 pscram-miR, pmiR-181a, and shRB1 cells (top) and a representative western blot of RB1 expression levels in FT237 pscram-miR, pmiR-181a, and shRB1 cells (bottom). b Micrographs showing anchorage independent growth of FT237 pscram-miR, pmiR-181a, and pshRB1 cells with quantification below. c Bar graph of the % Cell Cycle populations for the FT237 pscram-miR, pmiR-181a, and pshRB1 cells. d Pictures showing representative images of subcutaneous tumor formation at 25 weeks post injection for the FT237 pmiR-181a, and pshRB1 groups with insets depicting excised tumors (right) and graph of the number of injection sites that formed palpable tumors (left). Fisher’s exact test was used for statistical analysis. e Graph depicting tumor growth kinetics for the FT237 pmiR-181a and pshRB1 subcutaneous injection groups (left). Error bars represent ±SEM. Significance values are color coded to match the corresponding mouse injection group. Scatter plot showing total tumor burden for the FT237 pmiR-181a, and pshRB1 subcutaneous injection groups (right). N = 15 for the FT237 pmiR-181a group, N = 18 for the FT237 pshRB1 group. Mann–Whitney test was used for statistical analysis. f Graph showing the fold change of total number of CNV events for FT237 pscram-miR, pmiR-181a, and pshRB1 cells (inset) and the fold change of the number of events for each CNV subtype in FT237 pscram-miR, pmiR-181a, and pshRB1 cells. N = 2 for all cell lines. All data are representative of N = 3 independent experiments unless otherwise stated. The measure of center for the error bars is given as the mean value unless otherwise stated. The statistical test used for data analysis is the two-sided Student’s t test unless otherwise stated. Error bars indicate ± standard deviation unless otherwise stated. *p < 0.05, **p < 0.005, ***p < 0.0005. Full western blots shown in Supplementary Fig. 11.