Extended Data Fig. 3: Related to Fig. 4.
A) iMSC-related gene transcription in primary BM MSC with or without recombinant IL-1β. B) Gating for CD10+ neutrophils after coculture with (i)MSC, including representative plots after culture. C) C3AR, CD11b(act), and CD11c frequencies, or MFI of CD66b and CD45 on CD10+ neutrophils cultured alone (‘neutro only’), with rhIL-1β, or on ADSC-derived iMSC (n = 4; 3 experiments). D) BAFF protein in supernatant of neutrophils cultured alone, with IL-1β, or on ADSC-derived iMSC (3 donors; 2 experiments) E) BAFF or F) IL-1β protein in supernatant of (i)MSC cultured alone, neutrophils cultured alone, or neutrophils cultured on non-inflammatory ADSC-derived MSC or iMSC (4 donors; 2 experiments) G) BAFF H) or IL-1β protein in supernatant of neutrophils cultured alone, with different quantities of myeloma cells, or on ADSC-derived iMSC (4 donors; 2 experiments) I) Selected transcripts in unstimulated or IL-1β-stimulated ADSC. J) iMSC-signature enrichment plots (from DEGs of scRNA-seq in4), and selected Hallmark genesets for neutrophils cultured on iMSC or cultured alone. FDR, false discovery rate; Nom. P, nominal p-value; NES, normalized enrichment score K) C3AR, CD11b(act), and CD11c frequencies, or MFI of CD66b and CD45 on CD10+ neutrophils cultured alone, or on ADSC-derived MSC or iMSC (6 donors; 3 experiments). L) BAFF or M) IL-1β protein in supernatant of neutrophils cultured alone, or on ADSC-derived MSC, or iMSC (6 donors; 3 experiments) N) Volcano plot depicting DEGs of neutrophils cultured on ADSC-derived iMSC or cultured alone. Log2FoldChange cutoff, 1; adjusted p-value cutoff 10-4. Genes in red are related to activation O) Enrichment plots for MatNeu2 signature generated from data in Fig. 1 comparing neutrophils cultured on iMSC to cultured alone, or cultured on non-inflammatory MSC to neutrophils cultured alone. Data presented as mean ± SEM. Significance calculated in A and D using Mann-Whitney U test (two-tailed), in C, E, F, G, H, K, L and M using Wilcoxon Rank Sum test (two-tailed), in J and O using pre-ranked gene set enrichment analysis, and in I and N using Wald test (two-tailed) followed by Benjamini–Hochberg correction. *P ≤ 0.05, **P ≤ 0.01, ****P ≤ 0.0001, NS P > 0.05. Lines depict paired samples.
