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. Author manuscript; available in PMC: 2018 Mar 15.
Published in final edited form as: Clin Cancer Res. 2017 Jun 19;23(18):5446–5459. doi: 10.1158/1078-0432.CCR-17-0342

Figure 5.

Figure 5

Identification of chloride intracellular channel 4 (CLIC4) and RYBP as mediators of the effects of high score PSF. A) Gene expression signature of 50 most differentially expressed genes derived from MDA-MB-231 cells treated with 4 different high and 4 different low PSF samples. B) The 1000 most differentially expressed genes between high and low score treated samples were analyzed by Enrichr. GO Cellular Component Ontology identified focal adhesion, cell substrate adherens junction, and cell-substrate junction as the pathways most significantly associated with the differentially expressed genes upon high and low score treatment (*** p < 0.001). C) Western blot analysis of alpha smooth muscle actin (αSMA) as a marker of myofibroblast differentiation in primary human mammary fibroblasts treated with high or low score PSF. D) Quantification of αSMA in C. (Mean ± SEM)(p = 0.0411, High, n =4; Low, n = 4, Unpaired t-test). E–F) Immunohistochemistry of αSMA in mammary glands of nude mice treated with high or low score PSF. (scale bar = 100μm). G) The 1000 most differentially expressed genes between high and low score treated samples were analyzed by Enrichr. PANTHER Pathway identified the p53 pathway as the pathway most significantly associated with the differentially expressed genes upon high and low score treatment. (* p<0.05) H) Western blot analysis of p21 and p53 in mammary glands of nude mice treated with low and high score PSF. Actin was used as the loading control. I) Ductal histology analysis of mammary glands of p53515A/p53WT heterozygous mice untreated with PSF, or treated with high or low score PSF. Treated mice were treated bilaterally with high or low score PS. (n = 5 mice; n = 10 mammary glands per high or low score PSF treatment). J) H&E of a representative mammary gland from p53515A/p53WT heterozygous female mouse treated with low score PSF (scale bar = 100μm). K) H&E of a representative tumor in the mammary gland of a p53515A/p53WT heterozygous female mouse treated with high score PSF (scale bar = 100μm). L) Western blot analysis of p21 and p53 in the mammary gland of p53515A/p53WT heterozygous female mice treated with low and high score PSF.