Fig. 6

p66ShcA increases breast cancer cell dissemination into the bloodstream and subsequent lung colonization. a Number of circulating tumor cells (CTC) in mice bearing parental, p66-CR (VC), p66-CR (WT), and p66-CR (S36A) mammary tumors normalized both to tumor volume at necropsy and the volume of blood collected. The data is shown as average number of CTCs/mm³ tumor/ml blood ± SEM and is representative of 8 mice per group. b Percentage of surviving cells following matrix detachment (anoikis resistance) was determined for 4T1-537 parental, p66-CR (VC), p66-CR (WT), and p66-CR (S36A) cells. The percentage of surviving cells was normalized to that observed for adherent cells for each cell line. The data is representative of five replicates from three independent experiments. c Number of breast tumor cells present in the lung 1 h following tail vein injection. Breast cancer cells were labeled with Cell Tracker Red CMPTX dye and visualized in the lungs by fluorescent microscopy. The data is shown as average # cells per field of view ± SEM. For each cell line, the data is representative of 5 mice and 5 fields of view per mouse. Representative fluorescent images are shown. d Breast cancer cells were labeled and visualized as in panel b. The data is shown as the fold decrease in cell number 24 h post tail vein injection relative to 1 h ± SEM. The data is representative of 5 mice and 5 fields of view per mouse. Representative fluorescent images are shown. For panels a–d, statistical analysis was performed using a one-way ANOVA with a Tukey’s multiple comparison test (*P < 0.05; **P < 0.01; ***P < 0.001)