Fig. 3: ABHD11 inhibition activates SREBP signalling to drive oxysterol synthesis.

a Differential expression analysis by RNA-Seq in CD4+ effector T-cells (n = 4). Blue and red data points represent downregulated and upregulated genes, respectively. Transcripts with an adjusted p-value < 0.05 were considered differentially expressed. b Pathway enrichment analysis based on differentially-expressed genes (n = 4). Top 10 enriched pathways are shown. c Transcription factor enrichment analysis based on differentially-expressed genes (n = 4). The lower the “Average Rank” value, the more enriched the transcription factor activity. Top 10 enriched transcription factors are shown. d GSEA enrichment plot for Sterol Biosynthetic Process (n = 4). e Total intracellular sterol levels in CD4+ T-cells (n = 5). f Intracellular non-oxygenated sterol levels in CD4+ T-cells (n = 5). g Intracellular oxysterol levels in CD4+ T-cells (n = 5). h Intracellular levels of selected oxysterols in CD4+ T-cells. Metabolites include: 4β-hydroxycholesterol, 7α-hydroxycholesterol, 24-hydroxycholesterol, 25-hydroxycholesterol, 27-hydroxycholesterol, 24,25-epoxycholesterol (n = 5). Heatmap represented as Log₂(fold-change) versus vehicle control. i Expression of LXR-associated genes in CD4+ T-cells (n = 4). Heatmap represented as individual gene Z-scores. j IFNγ and IL-2 production by CD4+ T-cells, activated in the presence and absence of ML-226 or GW3965 (n = 6). For p < 0.0001, p = 0.00004089. k IFNγ and IL-2 production by CD4+ T-cells, activated in the presence and absence of ML-226 or oxysterols 27-hydroxycholesterol and 24,25-epoxycholesterol (n = 4). l IFNγ and IL-2 production by CD4+ T-cells, activated in the presence and absence of ML-226 and GSK2033 (n = 6). All experiments were carried out using human samples. CD4+ T-cells were activated with α-CD3 (2 μg/ml) and α-CD28 (20 μg/ml) for 24 h, in the presence and absence of ML-226, unless otherwise stated. Data are expressed as either: mean, with paired dots representing data from distinct biological replicates; or mean ± SEM. Statistical tests used: differential expression analysis via eBayesian fit of trimmed mean of M-values (a), two-tailed paired t-test (e–h), two-tailed one-sample t-test (j–l). Source data are provided as a Source Data file.