Fig. 5

REST-null GBM cells can rescue their growth through lipid metabolism rewiring. A Left, Western blotting confirms lack of REST protein in D4 cells. Proliferation of WT and D4 cells was examined by counting cells every 24 h. Shown is one representative replicate and quantification of PDT (right) based on three to four independent experiments (mean ± SD). Statistical comparison vs T98G control was performed using ANOVA with post hoc tests. Individual data values are provided in Additional File 10C. B Venn diagram showing overlap between upregulated DEGs in “slow” clones and “fast” D4 cells. C Gene ontology categories significantly enriched among D4-specific upregulated DEGs. D Gene network for “Fatty acid metabolic process” (GO:0006631) as a top-significant category from C. Network was built using STRING v.12 database. E Gene expression of ACSL1 and ACSL3 in T98G WT, “slow”, and “fast” REST-KO cells. Statistical difference between groups was tested using t-test. Shown are mean ± SEM from three biological replicates. Individual data values are provided in Additional File 10L. F Proliferation of D4 cells, in the presence of DMSO or pan-ACSL inhibitor Triacsin C (500 nM) in the media, was examined by counting cells every 24 h. Shown is one representative replicate and quantification of PDT based on three independent experiments. Statistical difference between groups was tested using two-tailed paired t-test. Individual data values are provided in Additional File 10 M. G Proliferation of D4 cells in the presence of Triacsin C (500 nM) does not differ from that of “slow” REST-KO clones. Comparison was performed using ANOVA with post hoc tests. Individual data values are provided in Additional File 10N. H Sensitivity of T98G WT and REST-KO cells to pan-ACSL inhibitor Triacsin C (72 h). Shown are LD50s with 95% confidence intervals calculated from three to four independent biological replicates. LD50s were compared vs T98G control using ratio test from “drc” R package. I Effects of GR-28 lead (4 μM for 18 h) on expression of select lipid metabolism genes from the network (D). Shown are fold changes (FC) vs DMSO derived from four independent biological replicates. Dashed line indicates FC = 1.5. Individual data values are provided in Additional File 10O. **p < 0.01; *p < 0.05; ns—not significant