Extended Data Fig. 6: Mitogenic tumor cues can elicit NRF2 activation in BM and TME to support myeloid expansion.

a, Frequency of HO-1⁺, Arg1⁺, MHCII⁺, and CD86⁺ BMDMs after culture with >3 kDa KP CM protein retentate fraction, and <3 kDa KP CM deproteinated fraction. n = 3 per group. b, Frequency of HO-1⁺, Arg1⁺, MHCII⁺, and CD86⁺ BMDMs after culture with heat-inactivated KP CM. N = 4 per group. c, Protein abundance of indicated analytes in tumor-associated BM sera, represented as relative fold-change over naïve sera. n = 2 per condition, one experiment. d–e, Analytes upregulated in BM sera (d) and blood sera (e) of tumor-bearing mice compared to naïve mice as assessed by O-link, with relative concentrations of GM-CSF and IL-6 (box). n = 3 per group, one experiment. f, Comparison of analyte concentration from BM sera and blood sera, data from (d) and (e). g, Ex vivo progenitor expansion assay, with frequency of ki67⁺ and HO-1⁺ myeloid cells upon culturing BM Kit⁺ cells with indicated growth factors for 2 days. n = 3 per group. h, Relative expansion (left, n = 4) and frequency of ki67⁺, HO-1⁺, and Arg1⁺ myeloid cells (right, n = 6) upon culturing progenitors with KP CM or indicated growth factors for 2 days. i–j, Relative expansion of myeloid cells with frequency of ki67⁺, HO-1⁺, and Arg1⁺ myeloid cells (i) and median fluorescence intensity (MFI) for CellROX and MitoSOX (j) after culturing progenitors with KP CM and indicated depletion antibodies for 2 days. n = 3 per group. k-l, Relative expansion of myeloid cells with frequency of ki67⁺, HO-1⁺, and NRF2⁺ myeloid cells (k) and MFI for CellROX and MitoSOX (l) after culturing progenitors with KP CM and ML385 for 2 days. n = 3 per group. m–n, Representative histogram and MFI quantification for phospho-p38 MAPK in myeloid progenitors treated with KP CM and indicated neutralizing antibodies (m, n = 3 per group) or KP CM and ML385 (n, n = 4 per group). o–p, Relative expansion of myeloid cells with frequency of ki67⁺, HO-1⁺, and Arg1⁺ myeloid cells (o) and MFI for CellROX and MitoSOX (p) after culturing BM progenitors with indicated growth factors in the presence of SB203580 for 2 days. n = 3 per group. q, Schematic for reactive oxygen species (ROS)-burden dependent poising of NRF2 pathways in growth factor-induced myeloid progenitor expansion in BM which enables NRF2 activation within TME. Individual data points with bar denoting mean, representative of two independent experiments (a,g,h,i–p). Statistics computed by one-way ANOVA with Tukey’s multiple comparison (a,b), Dunnett’s multiple comparison (g)–(p), hypergeometric test for O-link (d,e).

Source Data