Fig. 5: ACBP^KD and FAO inhibition downregulate invasion-related genes.

A Top: Venn diagram of differentially expressed genes in LN229 and NCH421K ACBP^KD cells compared to control cells (calculated with DESeq2, n = 5 and 4 samples per condition for LN229 and NCH421K, respectively, adjusted p-value <0.01). Bottom: List of common genes differentially expressed in ACBP^KD cells (both cell types) with color-coded log2 Fold changes. B LN229 Control or ACBP^KD cells were treated with either vehicle or Etomoxir (40 μM) for 5 days. Left: qRT-PCR data showing Itgb1 fold expression changes (mean ± SEM, n = 3 samples per group, one-way ANOVA with adjusted p-value for multiple comparisons *p < 0.05, **p < 0.01). Right: Immunostainings showing ITGB1 protein expression in each condition (ITGB1 channel was acquired with exactly the same microscopy parameters in all cases). Scale bars: 20 μM.C LN229 cells were transfected with either Control, Acbp or Itgb1 siRNAs and plated at confluency for transwell assays. Pictures show representative images of ITGB1 immunostainings in each condition (ITGB1 channel was acquired with exactly the same microscopy parameters in all cases). Scale bars: 20 μM. Transwell migration assay results for Control, Acbp or Itgb1 knockdown conditions (mean ± SEM, n = 3 samples per group, one-way ANOVA with adjusted p-value for multiple comparisons). D LN229 cells were transfected with either Control or Acbp siRNAs and transduced with either Control or ITGB1-overexpressing lentivirus. Cells from each group were plated at confluency for transwell assays. Pictures show representative bright field images of the upper (membrane) chamber and DAPI-stained migrating cells in the lower chambers, 4 days after plating. Scale bars: 50 μM (top) and 100 μM (bottom). Transwell migration assay results for each group (mean ± SEM, n = 4 samples per group, one-way ANOVA with Welch’s correction, adjusted p-value for multiple comparisons *p < 0.05, **p < 0.01).