Figure 4 from SREBP-Dependent Regulation of Lipid Homeostasis Is Required for Progression and Growth of Pancreatic Ductal Adenocarcinoma

SCAP is required for PDAC cell growth under low-serum and low-lipid conditions. A, Pa03c cells were grown for 16 hours in a medium containing 10% FBS or 0.5% FBS. mRNA expression was determined using microarray, and differentially expressed genes were identified (Supplementary Table S4). Bioinformatic gene set enrichment analysis was performed against four common pathway datasets. Results are shown using a normalized enrichment score (NES) of >2.0. B, Wild-type (WT) and SCAP KO Pa03c cells were grown for 7 days in a medium containing 10% FBS or 1% FBS in the absence or presence of lipid supplements (1 mmol/L mevalonate, 5 μg/mL cholesterol in ethanol, 20 μmol/L sodium oleate, and 50 μg/mL LDL). Media were changed every 3 days. Plates were stained with crystal violet as a measure of cell proliferation. Quantification of crystal violet staining is shown (n = 3 per group). For each cell line, growth was normalized to the 10% FBS condition. Statistical significance was determined using two-way ANOVA and Tukey HSD test. P values are indicated: *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001, ns, not significant. Error bar denotes SD. C, Cell growth assay of PDAC cell lines. WT, SCAP KO, and SCAP KO rescued cell lines were cultured in either 10% FBS supplemented with cholesterol (5 μg/mL), mevalonate (1 mmol/L) and oleate–albumin (20 μmol/L), or 10% LPDS with no additions for 7 days. Plates were stained with crystal violet. KEGG, Kyoto Encyclopedia of Genes and Genomes.