Fig. 3: DGKZ promotes epithelial–mesenchymal transition via the transforming growth factor β signaling pathway.

A Signaling pathways enriched based on the KEGG pathway analysis with the RNA sequencing data from DGKZ-upregulated BT-549 cells. B GSEA indicated that epithelial–mesenchymal transition and the transforming growth factor β signaling pathway were upregulated in DGKZ-overexpressing cells. C Protein levels of the EMT markers FN-1, E-cadherin, N-cadherin, and Snail and the TGFβ pathway markers p-Smad3, TGFβR1, TGFβR2, Smad3, and Smad4 in cell lines after DGKZ overexpression or downregulation based on western blot. D RNA expression of the TGFβ pathway markers TGFβ1, TGFβR1, TGFβR2, Smad3, and Smad4 in MDA-MB-231LM2 cells after DGKZ was downregulated based on real-time PCR. E F-actin staining of DGKZ-overexpressing MDA-MB-231 and Hs-578T cells or DGKZ-knockout MDA-MB-231LM2 and MDA-MB-231HM cells based on immunofluorescence. F Transwell assay of Hs-578T cells treated with the TGFβ receptor inhibitor LY2109761 (10 μM) for 48 h. G Transwell assay of MDA-MB-231LM2 cells treated with the pathway agonist TGFβ-1 (10 ng/l) for 48 h. H Western blot analysis of MDA-MB-231 and Hs-578T cells overexpressing DGKZ treated with LY2109761 (10 μM) for 48 h. I Western blot analysis of MDA-MB-231LM2 cells with stable knockout of DGKZ treated with TGFβ-1 (10 ng/l) for 48 h. J Western blot analysis of MDA-MB-231LM2 and MDA-MB-231HM cells downregulating DGKZ treated with cycloheximide (50 µM) for 24 h. The data represent the mean ± SD of three independent experiments. NS, no significance; Bars, ±SD; *p < 0.05.