Extended Data Fig. 6: Sapienate and cis-8-octadecenoate are used in membranes.
From: Evidence for an alternative fatty acid desaturation pathway increasing cancer plasticity
a–d, Heat map representing abundance changes of phosphatidylcholine (a), phosphatidylethanolamine (b), phosphatidylserine (c) and phosphatidylinositol (d) species in control and FADS2 knockdown HUH7 and A549 cells, relative to control. HUH7, control, n = 3; shFADS2-1, n = 4; shFADS2-2, n = 5; A549, n = 5. Only significant differences are depicted as fold change (log2(change with shRNA versus control)). X, blank or excluded values. Phospholipid species carrying sapienate or palmitoleate are depicted in bold red, and are listed in Supplementary Table 1e. Two-way ANOVA with Dunnett’s multiple comparisons. e, Relative distribution of phospholipid species in HUH7 (n = 2) and A549 (n = 5) cells with non-targeting shRNA (control). f, Membrane fluidity based on the ordered-to-disordered lipid ratio in HUH7 and A549 with a non-targeting shRNA (control, black) or one of two different shRNAs targeting FADS2 (brown (shFADS2-1) and orange (shFADS2-2)), normalized to control (n = 4). The higher the ordered-to-disordered ratio, the more saturated lipids are present in the membrane. One-way ANOVA with Dunnett’s multiple comparisons. g, Lipid peroxidation sensitivity via MDA assay in HUH7 with a non-targeting shRNA (control; black) or one of two different shRNAs targeting FADS2 (brown (shFADS2-1) and orange (shFADS2-2)) normalized to control (n = 3). Cells were treated with vehicle or 5 μM RSL3, which inhibits glutathione peroxidase 4 and induces lipid peroxidation. Two-way ANOVA with Sidak’s multiple comparisons. Cancer cell experiments were performed in low FBS DMEM (1%, HUH7; 0.5%, all other cancer cells) with a treatment of 72 h. Data are presented as mean ± s.d. from biologically independent samples.
