Fig. 2

DLAT silencing significantly reduces lipid content in OC cells and impairs OC cell proliferation and metastasis in vitro. (A) GSEA analysis of TCGA ovarian cancer dataset showing positive correlation between DLAT expression and ‘Cell Cycle’ and ‘Sphingolipid Metabolism’ gene signatures. (B) qRT-PCR and western blot analysis were used to confirm the transfection efficiency of two specific DLAT shRNAs (sh-DLAT#1 and sh-DLAT#2) in SKOV3 or OVCAR3 cells. (C-J) SKOV3 or OVCAR3 cells were transduced using sh-NC, sh-DLAT#1, or sh-DLAT#2 lentiviruses. Cells that were not transduced with lentivirus were used as controls. (C) Nile Red staining was used to measure cellular neutral lipids. Nuclei were stained with Hoechst. Scale bar = 50 μm. (D) Triglyceride levels were measured using a commercially available kit. Values were normalized to cellular proteins. Data are presented as mean ± SD and are representative of three independent experiments. (E) Cellular cholesterol content was measured using a commercially available kit. (F) Cell proliferation was assessed using the CCK-8 assay. (G) The EdU assay was used to assess cellular proliferation. Scale bar = 50 μm. (H) The colony formation assay was used to assess colony formation. (I) Transwell migration and invasion assays were used to assess the migratory and invasive abilities of cells. (J) The wound-healing migration assay was used to assess the effects of DLAT knockdown on cellular migration. Scale bar = 100 μm. Data are presented as the mean ± SD, n = 3. Statistical significance was determined by comparing each experimental group to the sh-NC control group. ***P < 0.001, **P < 0.01, and *P < 0.05