Extended Data Fig. 7: NRF2 signaling in tumor-infiltrating monocytic lineage promotes immunosuppressive program persistence in TME.
From: Myeloid progenitor dysregulation fuels immunosuppressive macrophages in tumours
a, Gene expression in KP conditioned media (CM)-exposed BMDMs as measured by reverse transcription quantitative PCR (RT–qPCR). Relative values depicted as fold-change compared to control media, after normalization to Hprt expression. n = 3 replicates, representative of two independent experiments. b, Gene expression in KP CM-exposed BMDMs treated with ML385, as measured by RT–qPCR. Relative values depicted as fold-change compared to BMDMs treated with vehicle DMSO, after normalization to Hprt expression. n = 3 replicates, representative of two independent experiments. c, Relative MFI quantification of CellROX, LiperFluo, and MitoSOX in Nfe2l2TKO (KO) or control (WT) BMDMs exposed to KP CM. n = 3 per group. d, Representative cytometry plots for Annexin-V and Propidium iodide (PI) in Nfe2l2TKO (KO) or control (WT) BMDMs exposed to KP CM, with relative frequency of apoptotic cells. n = 3 per group. e, Relative frequency of apoptotic cells in Nfe2l2TKO (KO) or WT BMDMs without any tumor conditioning. n = 3 per group. f–g, Number of LT-HSCs and myeloid progenitors in BM (f) and myeloid cells in blood (g) of tumor-free Nfe2l2ΔMs4a3 mice (n = 7) or Nfe2l2fl/fl controls (n = 5). h, MFI for indicated phenotypic markers in blood-circulating Ly6Chi monocytes from tumor-free Nfe2l2ΔMs4a3 mice (n = 5) or Nfe2l2fl/fl controls (n = 4). i, Fluorescent bead phagocytosis assay using tumor-free Nfe2l2ΔMs4a3 or Nfe2l2fl/fl BM Ly6Chi monocytes at different ratio of cells to beads. n = 4 per group. j, frequency of NRF2+ CD45.2+ donor-derived mo-macs derived from GMPs in Brusatol-treated tumor-bearing mice or N-acetyl cysteine-treated tumor-naïve mice transferred into KP tumor-bearing CD45.1 hosts, four days post transfer. (left to right, n = 3,4,4,3). Individual data points with bar denoting mean, representative of two independent experiments (a,b,f–j) or three independent experiments (c,d,e). Statistics computed by unpaired two-tailed Student’s t-test (c–h), two-way ANOVA with Fisher’s LSD test (a,b) or Sidak’s multiple comparison (i).
