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Fig. 2 | BMC Biology

Fig. 2

From: The Z-cad dual fluorescent sensor detects dynamic changes between the epithelial and mesenchymal cellular states

Fig. 2

The Z-cad dual sensor detects MET in claudin-low human and mouse breast cancers. a (Left) The d2GFP-Z1 3′ UTR sensor constitutively expresses destabilized GFP (d2GFP) under control of constitutively active ubiquitin C promoter (UbC). The human ZEB1 3′ UTR containing eight miR-200 binding sites was cloned downstream of d2GFP. (Right) The Ecad-RFP sensor contains about 1370 bp of the human E-cadherin promoter regulating dsRED (RFP) expression. Three E-boxes proximal to transcription start site are indicated. Co-transduction of both sensors yields cells containing the Z-cad dual sensor. b Flow cytometric analysis of human MDA-MB-231 breast cancer cells, mouse T11 breast cancer cells, and human BLSL12 PDX-derived cells containing the Z-cad sensor. miR-200c/141 was induced with 2 μg/mL doxycycline for 4 days and flow cytometry was performed. Epithelial (MET)- and mesenchymal (EMT)-enriched populations are as indicated (n = 3 biological replicates per group). c Fluorescent confocal microscopy of MDA-MB-231 cells imaged at indicated time points during induction (+DOX) of miR-200c. –DOX is non-induced control. Scale bar = 50 μm

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