2–2.0 × 106 cells per host). Tumors and blood of the recipient animals were analyzed by flow cytometry for the presence of grafted cells 24, 72, and 120 h after the transfer. Doxorubicin (Ebewe Pharma, Kundl, Austria) was injected into tumor-bearing animals in a single dose of 10 mg/kg i.p., control animals were injected with 150 μL PBS [4]. Mice were sacrificed 24 and 48 h thereafter. The CSF1R inhibitor GW2580 (LC Laboratories, Woburn, MA) was given orally in daily dose of 180 mg/kg as described [22] for 48, 96 h or 2 weeks. Control mice were treated with vehicle alone. Animals were additionally injected with www.selleckchem.com/products/AG-014699.html 1 mg

BrdU 3 h before necropsy. The treatments were initiated 1 week after tumor recognition. Cellularity of organs was analyzed by flow 26s Proteasome structure cytometry. Z scores were calculated from log2 gene expression values within each patient cohort and correlated together with tumor stage and ER-status as covariates using linear-model multiple regression. In case of the pooled data,

the random effect of particular study was included in the regression model. HR and their significance for overall patient’s survival were obtained with Cox regression. Statistical significance of the remaining data was assessed with two-tailed Student’s t-test, one-way or two-way ANOVA with Bonferroni post hoc test as appropriate. Significances for analyses with more than two factors were assessed with linear-model multiple regression and Bonferroni-corrected t-tests as post hoc tests. Probability values presented in figures refer to the results of the post hoc tests and the significances for the main Nutlin-3 factors determined by ANOVA or multiple regression. The difference was considered significant when p < 0.05. For 0.05 ≤ p < 0.10, the probability value was also presented. If not stated otherwise, each symbol on the plot indicates one individual animal and is represented together with mean ± SEM from

n individual animals. Statistical analyses were performed with GraphPad Prism (GraphPad Software, La Jolla, CA) and R Platform software. This work was supported by grants from the Integrated Center for Research and Therapy (IFTZ) of Innsbruck Medical University (W.D.), the Austrian Cancer Society/Tirol and the doctorate program MCBO funded by the Austrian Science Fund FWF (P.T. and S.D). We would like to thank Pornpimol Charoentong, MSc, for help with data analysis and Dr. Michael Haffner, Dr. Andreas Villunger, Dr. Gerrit-Jan Wiegers, and Dr. Patrizia Stoitzner for fruitful discussions. We acknowledge Anto Nogalo for excellent technical assistance, Dr. Ernst Werner for providing primers for qRT-PCR analysis, Dr. Gerold Untergasser for providing CD31 antibody, and Dr. Roswitha Sgonc for providing access to cryocut device. The authors declare no financial or commercial conflict of interest.