Fig. 4: Expression of RELB in TSKI4 cells induces EMT.
From: Coordinated regulation of Rel expression by MAP3K4, CBP, and HDAC6 controls phenotypic switching
a RELB protein levels were measured in TSWT cells, TSKI4 cells, and TSKI4 cells expressing RELB (TSKI4RB) cells. Western blots of nuclear lysates are representative of n = 2 biologically independent experiments. b RELB expression induces a mesenchymal morphology in TSKI4R cells. Representative phase microscopy images for n = 3 biologically independent experiments are shown. Scale bar represents 400 μm. Arrows show the area of enlarged insets. c RELB expression in TSKI4 cells results in loss of epithelial marker expression. Transcripts were measured using qPCR. The data normalized to Rps11 are expressed as fold-change relative to TSWT cells and are the mean ± SEM of n = 3 (Cdh1) or n = 5 (Cldn6) biologically independent experiments. d–f Increased transcripts of mesenchymal markers (d, e) and EMT-inducing TFs (f) upon RELB expression. Transcripts were measured using qPCR. The data normalized to Rps11 are expressed as fold-change relative to TSWT cells and are the mean ± SEM of n = 3 biologically independent experiments. g Invasion through growth factor reduced Matrigel was increased in TSKI4RB cells relative to TSKI4 cells. Data show the mean ± range of n = 2 biologically independent experiments performed in triplicate. h Barrier formation shown as the fold change in fluorescence of diffused dye relative to TSKI4 cells. Data are expressed as the mean ± SEM of n = 3 biologically independent experiments. *p-value < 0.05; ***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.
