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. 2016 Dec 12;213(13):2967–2988. doi: 10.1084/jem.20151665

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© 2016 Chan et al.

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Figure 3. — MTD chemotherapy induces chronic ELR⁺ chemokine expression in CAFs. (A) Heat map depicting hierarchical clustering of 3,006 differentially expressed genes (DEGs) detected in vehicle (Veh)-, MTD-doxorubicin–, or MTD-paclitaxel–treated BCAF-011 CAFs. The map depicts high (yellow) and low (blue) relative levels of medium-centered gene expression in log space (n = 2 at each time point). (B) Network map of enriched functional gene categories in the 2,046 late-response genes (i.e., differentially expressed genes at day 7 and/or day 14 after initiation of the treatment) detected in the chemotherapy-treated CAFs. GO-BP, gene ontology biological process; GO-CC, gene ontology cellular component; SPK, SwissProt keywords. (C) Relative transcript levels of the ELR⁺ chemokines CXCL1, CXCL2, CXCL5, and CXCL6 in vehicle-, MTD-doxorubicin–, or MTD-paclitaxel–treated CAFs. Data from two independent experiments (n = 3 in each group; ***, P < 0.001) are shown. (D) The amounts of the secreted ELR⁺ chemokines detected in the conditioned medium from vehicle- and MTD-doxorubicin– or MTD-paclitaxel–treated CAFs. Data from two independent experiments (n = 3 in each group; *, P < 0.05; ***, P < 0.001) are shown. ND, not detected. (E) HistoGene-stained breast cancer tissue sections before (left) and after (right) LCM. The microdissected samples (top right) containing mainly (95%) stromal fibroblasts (yellow arrows), and those containing cancer cells were collected from the same tissue sections for subsequent RNA isolation. White arrows indicate carcinoma cells. Bars, 50 µm. (F) Relative transcript levels of the ELR⁺ chemokines measured in laser-captured tissue excised from the CAF-enriched tumor stroma (left) or the adjacent carcinoma cell–enriched region of neoadjuvant-treated or untreated human breast cancer tissues (right). At least six dissections were performed from different tumor areas from each sample. Data from three independent pairs of tumor samples (***, P < 0.001 vs. untreated tumors) are shown. Data are mean ± SEM; Student’s t test.