Fig. 1: Senescent B-lymphoma cells exhibit phenotypic and functional myeloid plasticity.
From: Senescence-associated lineage-aberrant plasticity evokes T-cell-mediated tumor control
a Induced gene-regulatory network47 of upregulated genes from the Eμ-myc;bcl2 therapy-induced senescence (TIS) model39. Red boxes represent interconnected regulatory hubs consisting of AP-1, PU.1 and C/EBPβ transcription factors (TF); blue box marks a TIS-related NF-κB hub. b Capillary-based immunoassay of hematopoietic TF in nuclear lysates of control;bcl2 (n = 8) and Suv39h1−;bcl2 (n = 6) lymphomas, 5-day exposed to adriamycin (ADR) or left untreated (UT) (left). TATA box-binding protein (TBP) as loading control. Representative digital blots of three lymphomas per genotype. Mean TF fold-induction values ± SEM compared to matched UT lymphomas (right). c Hierarchical clustering of top 100 TIS-upregulated genes as in a (93 measured and shown), for their expression in hematopoietic lineages from ‘Haemopedia’56. d tSNE clustering of scRNA-seq profiles from conditionally senescent (TIS, red dots) and non-senescent (non-TIS, blue dots) Suv39h1-inducible lymphomas16 (top left); specific analyses of B-cell transcripts, enrichment scores of PU.1 and dendritic cell (DC), adult tissue stem cell signature (ATSC) previously associated with senescence16,57, and Ncf2 expression as indicated. e Representative May-Gruenwald-Giemsa stain (n = 5 lymphomas per genotype); ANA-1 macrophages for comparison. Scale bars, 5 μm. f Flow cytometry-determined percentage of CD11b+ (left) and CD115+ (right) control;bcl2 (n = 21) and Suv39h1−;bcl2 (n = 17) lymphomas as in b. g Respiratory burst of control;bcl2 (n = 6) and Suv39h1−;bcl2 (n = 4) lymphomas 5 days after ADR or palbociclib (Palbo) treatment or left UT. Representative microscopy images (left) and quantification (right). Arrows indicate reduced nitroblue-tetrazolium (NBT) precipitates. Scale bars, 100 μm or 20 μm for inlays. ANA-1-macrophages (middle) with the percentage of reduced NBT+ cells ± SD (n = 3 technical replicates). f, g Bars indicate the mean percentage of positive cells ± SEM. Two-sided P-values by unpaired t-test on log2-transformed data (b); by paired t-test for UT vs. ADR comparisons of the same genotype and unpaired t-test for control vs. Suv39h1– comparisons (f). * two-sided P by non-parametric Friedman test comparing senescent to UT conditions (P = 0.006) or by two-way ANOVA assessing the contribution of Suv39h1– genotype to the data variance (P = 0.032) (g). Source data are provided in the Source Data file.
