Fig 2: CAF-derived NLRP3 inflammasome signalling facilitates tumour growth. a Scheme of experiments analysed in b–h. shNlrp3 AT3 cells were orthotopically co-injected with WT (Nlrp3+/+) or with Nlrp3 -/- NMFs (C57BL/6) into C57BL/6 mice. b Growth curves of AT3 tumours. n = 7 mice per group. c Tumour weights at termination of experiment. n = 6 and 7 mice per group (WT and Nlrp3-/-, respectively). d Flow cytometry analysis of Gr1+ cell infiltration into AT3 tumours co-injected with WT NMFs (n = 4) or with Nlrp3-/- NMFs (n = 3). Data presented are percentage of CD45+ cells, normalised to WT. e Representative images of staining with anti-Gr1 antibody in AT3 tumours. Scale bar, 100 µm. f Quantification of staining shown in d. Three fields of 3 mice/group were analysed; Data are presented as mean Gr1+ cells/field ± s.d; Mann–Whitney test. g, h Flow cytometry analysis of CD11b+Ly6Chigh Ly6G- and CD11b+Ly6G+Ly6C- cell infiltration into AT3 tumours co-injected with WT NMFs (n = 4) or with Nlrp3-/- NMFs (n = 3). Data presented are percentage of CD45+ cells, normalised to WT. i Scheme of experiments analysed in j–r. Met-1 cells were orthotopically co-injected with NMFs depleted of Nlrp3 or Il1b expression (shNlrp3, and shIl1b), or with control NMFs (shScramble). j Growth curve and k weight at endpoint of Met-1 tumours. n = 5, 10, 20, 19 tumours (TCs only, shScramble, shNlrp3, and shIl1b, respectively). l–n Flow cytometry analysis of Gr1+ cell infiltration into Met-1 tumours. n = 7, 14, and 16 per group (shScramble, shNlrp3, and shIl1b, respectively). Data presented are percentage of CD45+ CD11b+ cells, normalised to shScramble. o–r Met-1 tumours were analysed for the expression of selected genes using qRT-PCR. n = 6, 7, and 5–6 tumours per group (shScramble, shNlrp3, and shIl1b, respectively). Data are presented as fold change from shScramble. In b–d, g, h, j, k, data are presented as mean ± s.e.m; Welch’s t-test, *p < 0.05, **p < 0.005. In j–r data are presented as mean ± s.e.m; One-way analysis of variance test followed by Tukey’s multiple comparisons test. Data are representative of three independent biological experiments. Source data are provided as a Source Data file

Fig 3: Loss of CXCR4 Leads to Differentiation of Leukemia Cells(A) Representative histogram showing total ROS levels (measured with CellROX Deep Red) in c-Kit+ cells within GFP+ (sgRNA-expressing) cells harvested from the bone marrow of recipient mice (n = 5 for control sgRNA; n = 4 for Cxcr4 sgRNA_b) 14 days after transplantation.(B–D) Mean fluorescence intensity (MFI) of (B) phosphorylated p38 MAPK, (C) phosphorylated NF-?B, and (D) Gr-1 within GFP+ (sgRNA-expressing) leukemia cells that were harvested from the bone marrow of recipient mice (n = 3 for each group) 10 days after transplantation.(E) May-Grünwald-Giemsa-stained cytospin slides from representative sgRNA-expressing leukemia cells 13 days after transplantation. Scale bars represent 10 µm.Means and standard deviations are shown (*p < 0.05, **p < 0.01, ***p < 0.001). See also Figure S5.

Fig 4: Genetic KO of Usp15 Impairs HSC Function(A–E) Competitive transplantation of BM cells freshly isolated from Usp15+/+ or Usp15-/- mice.(A and B) Chimerism in peripheral blood (A) and contribution of BM cells to myeloid (Gr1+), B cell (CD19+) and T cell (CD3+) lineages in the blood (B) of recipients.(C) Representative FACS profiles (left) and numbers of donor-derived LSKs per million viable BM cells in recipients at 18 wpt.(D) Representative FACS profiles (left) and numbers of donor-derived HSCs (LSK/CD150+/CD48-) per million viable BM cells in recipients at 18 wpt.(E) Numbers of donor-derived myeloid committed progenitor populations (LKSs-, CMPs, GMPs, and MEPs) in recipients at 18 wpt.(F) FACS-sorted LSKs were plated after 8 days (second plating) in culture and monitored for growth. n = 4 wells per data point. Representative images at days 3 and 8 of the second plating are shown. Bar, 20 µm.*p = 0.05; **p = 0.01; ***p = 0.001. Error bars represent mean ± SEM (A–E); data represent one representative experiment out of three. See also Figure S5.

Fig 5: Induced proliferation of lin-Ter119-Mac-1-Gr-1-c-Kit+CD71(low/-) mouse bone marrow cells by over-expression of the transcription factor c-Myc. Shown are average measurements from 3 separate in vitro experiments where cells were modified by stable gene transfer of the TRE3G-cMyc vector, cultured in puromycin to eliminate non-modified cells (except wild type condition), then cultured with 2000 ng/ml dox for expression of the vector gene of interest or no dox as a control. Passage number and split ratio were used to determine fold expansion of total cells from the starting well and shown for 6 weeks of culture