Fig. 3: Induction of REL expression in mesenchymal-like TS^KI4 cells promotes an epithelial phenotype.

a Human REL transcript levels were measured in uninduced and Doxycycline (Dox) induced TS^KI4 cells transduced with lentiviral constructs expressing Tet-on-human REL (TS^KI4R) using qPCR and human specific REL primers. The data normalized to Rps11 are expressed as a fold-change relative to TS^KI4 cells and are the mean ± SEM of n = 4 biologically independent experiments. b REL protein expression was measured using Western blotting with anti-human specific REL antibody. Blots of whole-cell lysates are representative of n = 3 biologically independent experiments. c Dox induced REL expression restores an epithelial morphology in TS^KI4 cells re-expressing REL (TS^KI4R) cells. Representative phase microscopy images for n = 3 biologically independent experiments are shown. Scale bar represents 400 μm. d REL expression in TS^KI4 cells increased transcripts of epithelial markers. e–g Reduced mesenchymal markers (e, f) and EMT-inducing TFs (g) transcripts upon REL re-expression. d–g Transcripts were measured using qPCR. The data normalized to Rps11 are expressed as a fold-change relative to TS^WT cells and are the mean ± SEM. d n = 4 (Cldn6) and n = 3 (Krt8, Krt18) biologically independent experiments are shown. e, f n = 4 biologically independent experiments are shown. g n = 3 biologically independent experiments are shown. h Invasion through growth factor reduced Matrigel was decreased in Dox induced TS^KI4R cells relative to uninduced TS^KI4R cells. Data show the mean ± range of n = 2 biologically independent experiments performed in triplicate. i Barrier formation is increased in TS^KI4 cells after REL re-expression. Data show the fold change in fluorescence of diffused dye relative to TS^WT cells and are the mean ± SEM of n = 3 biologically independent experiments. *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001, ****p-value < 0.0001; Student’s t test. (n.s. not significant). Western blots show cropped images. Uncropped images are available in Supplementary Materials. See also Supplementary Figs. 2 and  3.