Figure 5.

Rank ectopic expression increases breast cancer aggressiveness

(A) Relative tumor growth and the percentage of lung metastasis in the indicated genotypes. Mean tumor volume ± SEM at each time point relative to their volume on the first day of detection and statistical t test values are shown. The percentage of females with the indicated number of lung metastasis and total number of mice is indicated.

(B) Number, size and representative pictures of secondary tumorspheres derived from PyMT^+/− and PyMT^+/−Rank^+/tg tumor cells. Each bar represents a pool of 3 independent tumors/experiments. Mean, SEM, and t test p values are shown.

(C) Metastasis initiating cell frequencies (with confidence intervals) calculated by ELDA, p- and chi-square values of PyMT^+/− and PyMT^+/−Rank^+/tg tumor cells are shown. Cells from 2 independent tumors per genotype were pooled for injections and metastasis were scored 8 weeks later.

(D) Frequency of indicated cells in the Lin⁻ (CD45⁻ CD31⁻) population found in PyMT^+/− and PyMT^+/−Rank^+/tg spontaneous tumors (n = 5–7) analyzed by FACS. Positive/negative and high/low populations were set according to cell populations in the normal MG. Mean, SEM, and t test p values are shown.

(E) Representative CK14 or CK5 (green) and CK8 (red) immunostaining in PyMT^+/− and PyMT^+/−Rank^+/tg spontaneous adenocarcinomas. Asterisks indicate magnifications (2×) to highlight CK14+/CK8+ cells. No CK5+/CK8+ cells were found.

(F) Quantification of CK8+, CK14+ and CK14+/CK8+ cells in spontaneous adenocarcinomas of PyMT^+/− and PyMT^+/−Rank^+/tg mice pretreated or not with navitoclax.

(G) Relative tumor growth curves in PyMT^+/−Rank^+/tg mice treated or not with navitoclax for 14 days. Mean tumor volume ± SEM at each time point relative to their volume on the first day of detection and statistical t test p values are shown. The number of mice is indicated.

(F and G) Navitoclax treatment started (at 4 weeks) before palpable lesions were detected. See also Figure S5.