Fig. 4: Suppressing FA uptake impairs NB cell survival.
From: MYCN-driven fatty acid uptake is a metabolic vulnerability in neuroblastoma
a Silencing SLC27A2 in LAN5 cells. Two shSLC27A2 GIPZ vectors tested, with empty GIPZ vector as control. Mean ± SEM (n = 3). b FA uptake in LAN5 shCTRL and shSLC27A2 cells. Cells stained with the FA analog BODIPY™ 558/568 C12 and quantified as CTCF by ImageJ2. Mean±SD (n = 3). c Cell growth in LAN5 shCTRL and shSLC27A2 cells. Mean ± SD (n = 3). d Clonogenic assay in LAN5 shCTRL and shSLC27A2 cells. Mean ± SD (n = 3); e–h LAN5 shCTRL and shSLC27A2 orthotopic xenograft model. e Tumor volumes at weeks 3 and 4 post-implantation. CTRL = 15; shSLC27A2 = 14; two-way ANOVA with Sidak’s multiple comparisons test. f Tumor weights at week 5 post-implantation. Mean ± SEM (CTRL = 13, shSLC27A2 = 12); two-sided unpaired Mann–Whitney test. g Oil Red O staining of intratumoral lipids. Mean ± SEM (CTRL = 6, shSLC27A2 = 8); two-sided unpaired Mann–Whitney test. h Lipidomics profiling of shCTRL and shSLC27A2 tumors (n = 8 each). Lipids (FDR < 0.25, absolute FC > 2) are shown in the heatmap (color scaled by Z-score: red = upregulated; blue = downregulated). MGDG monogalactosyldiacylglycerol, DGDG digalactosyldiacylglycerol, dhCER dihydroceramides, CL cardiolipin, SM sphingomyelin, PA phosphatidic acid, lyso.PC lysophosphatidylcholine, lyso.PE lysophosphatidylethanolamine, plasmenyl.PC plasmenylphosphatidylcholine, all other abbreviations consistent with Fig. 1e. i FA uptake following CB5 treatment in LAN5 and SHEP cells (0–15 µM, 5 min). Cells stained with BODIPY™ 500/510 C1, C12 and quantified as CTCF by ImageJ2. Mean ± SD (n = 3). j Caspase 3/7 activity of NB and normal cells after CB5 treatment (0–10 µM, 24 h). Mean ± SD (n = 3). k Apoptosis, p53/p21, and MYCN/c-MYC protein expression in LAN5, IMR32, and SHEP with CB5 (0–20 µM, 16–24 h). VP16 (10 µM, 24 h) = positive CTRL. Representative blots from three independent experiments are shown. FC fold change, Arb. Unit arbitrary unit. a, b, d, and i, one-way ANOVA with Dunnett’s multiple comparisons test; c and j, two-way ANOVA with Dunnett’s multiple comparisons test. Source data are provided in the Source Data file.
