Fig. 5.
Early loss of Fbxw7 results in dysregulated c-Myc, E2F, and Notch-ICD target expression. A. Mammary epithelial cells were isolated from two control (Cre-) and two Fbxw7−/− (Cre+) mice during the third week of pregnancy. PCR analysis for the excision of exons 5–6 of Fbxw7 was performed as in Fig. 1A. L = molecular weight ladder. B-E. Single cells were isolated from the populations shown in A using the Fluidigm C1 system and qPCR assays were performed using a Biomark HD (Fluidigm). Target amplification was performed with Delta Gene assays (primer sets) from Fluidigm for (B) c-Jun, (C) c-Myc, (D) E2F, and (E) Notch1-ICD target genes of interest. Lanes marked with + indicate Fbxw7+/+ control cells and lanes marked with - indicate Fbxw7−/− cells. Statistical analysis was performed using the Fluidigm Real-Time PCR Analysis software. Data are expressed as Log2Expression normalized to RNA spike-in controls. Significance was measured using the Mann-Whitney test with a p value cutoff of p < 0.05. *p < 0.05, **p < 0.01, ***p < 0.001. Arrows indicate genes exhibiting a distinct bimodal distribution. See Supplementary Tables 2–5 for data included in these analyses.