Figure 4. GALNT3 Re-expression in TS^KI4 Cells Restores an Epithelial Phenotype.

(A) Increased Galnt3 transcripts in TS^KI4 cells expressing human GALNT3. Transcripts were measured using qPCR in TS^WT and TS^KI4 cells infected with a control lentiviral construct (C) or TS^KI4 cells infected with a lentiviral construct expressing human Galnt3 (TS^KI4Gab1 and TS^KI4Gab2).

(B and C) Re-expression of GALNT3 in TS^KI4Gab1 and TS^KI4Gab2 cells was measured by western blotting.

(B) Blots are representative of three independent experiments.

(C) Densitometry was used to quantify three independent experiments.

(D) GALNT3 re-expression restores an epithelial morphology. Representative phase microscopy images from three independent experiments are shown. Black bar represents 100 μm.

(E) Increased transcripts of epithelial markers with re-expression of GALNT3.

(F and G) Western blots of E-cadherin (F) and Claudin-6 (G) are representative of three independent experiments.

(H) GALNT3 re-expression promotes barrier formation. Diffusion is expressed as a fold-change in fluorescence relative to TS^WT cells. Data shown are the mean ± SEM of three independent experiments.

(I and J) Re-expression of GALNT3 reduces mesenchymal markers (I) and EMT-inducing transcription factors (J).

(K) Re-expression of GALNT3 in TS^KI4 cells reduces invasiveness through growth-factor-reduced Matrigel. Data show cells per 10× field and are the mean ± range of two independent experiments performed in triplicate.

(A, E, I, and J) qPCR data normalized to Actb are expressed as a fold-change relative to TS^WT cells and are the mean ± SEM of three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001; Student’s t test.

See also Figure S3.