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Fig. 3 | BMC Biology

Fig. 3

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

Fig. 3

The Z-cad sensor identifies EMT induced by TGFβ1 in HMLER cells. a F-actin stress fibers increase during TGFβ1 compared to vehicle treatment (26 days) indicating EMT. Scale bar = 100 μm. b RNA analysis by qRT-PCR at indicated time points during vehicle or TGFβ1 treatment (n = 3 biological replicates per time point). Vehicle-treated values were set to 1.0 for each gene and analyzed using an unpaired Student’s t test. c Fluorescent confocal microscopy indicating a gain of GFP and loss of RFP at 14 days of TGFβ1 versus vehicle treatment. Mesenchymal-appearing bright GFP cells are indicated by arrows. Scale bar = 20 μm. d Flow cytometric analysis of HMLER cells containing Z-cad sensor showing an increase in cells displaying GFPhiRFPlow/neg EMT fluorescence signature during TGFβ1 treatment at the times indicated. e Quantitation of cells displaying the EMT signature in HMLER cells containing Z-cad dual sensor from d. Bar graphs depict the percentage of cells displaying the EMT fluorescence signature in TGFβ1- versus vehicle-treated groups (n = 3 biological replicates per time point). Data were analyzed using an unpaired Student’s t test. * p value < 0.05, ** p value < 0.01

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