Fig. 5: The adipogenic stimulation by IL-1β is limited to the early differentiation stage.

a RNA-seq of differentiating hASCs treated with IL-1β from start of adipogenic induction and collected on day 2, 5, and 9 of differentiation. Volcano plots show genes significantly regulated by IL-1β (adjusted p-value < 0.05, fold change > 1.5 and < –1.5) compared to vehicle, as analyzed by Wald test corrected for multiple testing using the Benjamini-Hochberg method. The five most significantly regulated genes on each day are labelled (n = 4 independent experiments). b Gene Set Enrichment Analysis of RNA-seq data described in (a). The 12 most highly enriched Hallmark gene sets (by normalized enrichment score, NES) in IL-1β-treated cells on each day are shown. Arrows indicate pathways directly related to adipogenesis or lipid handling. c Lipid droplet accumulation on day 9 of differentiation in hASCs treated with IL-1β at indicated differentiation stages (n = 4 independent experiments with ≥ 4 replicates (52 datapoints for vehicle and 16 for the rest)). d, e Adipogenesis- and adipocyte phenotype-related gene expression in hASCs treated with IL-1β on day 0 (d) or 7 (e) of differentiation and collected 48 h later (n = 3 independent experiments with 2 replicates). f IL1R1 expression, measured by Cap Analysis of Gene Expression (CAGE), of hASCs at indicated time points after adipogenic induction (n = 3 replicates). g, h Expression of pro-inflammatory genes in hASCs treated with IL-1β on day 0 (g) or 7 (h) of differentiation and collected 48 h later (n = 3 independent experiments with 2 replicates). Statistical analyses by one-way ANOVA and Dunnett’s multiple comparisons tests compared to vehicle (c), to IL-1β treatment days 0–5 (c), or to 0 h (f), or two-sided unpaired t-test (d, e, g, h). IL-1β: 10 ng/ml in all panels. Data are represented as box-and-whisker plots (line inside box = median; box limits = first and third quartiles; whisker ends = minima and maxima) or individual measurements and mean ± SEM. § denotes conditions with undetectable mRNA levels in all or some samples. Source data are provided as a Source Data File.