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. 2020 Dec 1;34(23-24):1697–1712. doi: 10.1101/gad.340604.120

Figure 2.

Figure 2.

DUSP11 alleviates the interferon response mediated by tumor–fibroblast cell interaction. (A) Schematic diagram of coculture assay (left), and RT-qPCR analysis (right) of IFNB1, MX1, and ISG15 mRNA induction normalized to GAPDH mRNA in MDA-MB-231 breast cancer (BrCa) cells and HFF fibroblast cells either in monoculture or coculture. Cells were cultured for 60–72 h and RNA lysates were collected for RT-qPCR analysis. RT-qPCR results are presented relative to monocultured MDA-MB-231 cells. (B) RT-qPCR analysis (left) of IFNB1 and ISG mRNA transcripts (MX1, ISG15, and IFIT1 mRNA) comparing fold change of cocultured cells silenced of DUSP11 (siD11 coculture) relative to control coculture (siNC coculture), and immunoblot analysis (right) assessing siRNA knockdown of DUSP11 in MDA-MB-231 and HFF cells treated with negative control siRNA (siNC) or siRNA targeting DUSP11 (siD11). (C) RT-qPCR analysis (left) of IFNB1 and MX1 mRNA normalized to GAPDH mRNA in cocultured MDA-MB-231 and HFF cells transduced with pLenti empty vector (vector) or pLenti-DUSP11-3xFLAG (D11), and immunoblot analysis (right) of endogenous and 3xFLAG-tagged DUSP11 in whole-cell lysates. RT-qPCR results are presented relative to those of monocultured MDA-MB-231 vector cells. Data are derived from n = 4 independent replicates in A and B and n = 3 independent replicates in C. In all panels, data are presented as mean ± SEM. (*) P < 0.05 (two-tailed Student's t-test).