Fig. 1

MDMX and MDM2 knockdown in MDA-MB-231 orthotopic transplants reduces CTCs. MDA-MB-231 cells with constitutive shmdm2, shmdmx, or mir30 shRNA-expressing vector were implanted into the mammary fat pads of 6-week-old female NSG mice. a Western blot analysis of MDM2, MDMX, and mtp53 protein levels from 50 μg of whole-cell lysates from 231.mir30.vector, 231.shmdm2, and 231.shmdmx cells (lanes 1, 2, and 3, respectively) prior to mammary fat pad implantation. Actin is shown as a loading control. b Box-and-whisker plot represents the numbers of CTCs per milliliter from 231.mir30.vector, 231.shmdm2, and 231.shmdmx cells engrafted into animals. The number of CTCs was determined by flow cytometry, and the total events were counted (gates were set by the GFP signal intensity and cell size). The number of CTCs per milliliter was obtained by dividing the number of positive events by blood volume from individual animals. The adjusted p value was obtained with two-tailed, two-sample t tests using a permutation test. c Representative fluorescence-activated cell sorting plots showing GFP-positive events in different mouse groups