Extended Data Fig. 10: VIMhigh macrophages promote the immune suppressive ability of Treg via IL-1β.
a, Violin plots showing the expression levels of indicated genes in VIMhigh and VIMlow macrophages. Two-sided Wilcox tests. The p.adj was calculated by Bonferroni correction. b, Spearman correlations (two-sided) between IL1B and the signature score of VIMhigh macrophages in three independent bulk-seq cohorts. 95% confidence intervals were shown. EHBH cohort, n = 237. HCCDB6, n = 445. HCCDB15, n = 400. c, Spearman correlations (two-sided) between IL1B and the signature score of VIMhigh macrophages in the indicated clusters in ST datasets. 95% confidence intervals were shown. d, Representative gating results showing the expression of IL-1β and Vimentin in indicated cells. e-g, Indicated mRNA level of genes related to macrophage differentiation in THP1 (e, n = 4), U937 (f, n = 4), and hMDM (g, n = 3). Two-sided unpaired t-test. h, The successful overexpression of VIM was checked by Western Blot and qRT-PCR in THP1 and U937 cells. Two-sided unpaired t-test. n = 3. i, The secretion level of IL-1β detected in indicated status of U937 Ctrl and VIMOE cells. Two-sided unpaired t-test. n = 3. j, The secretion level of Vimentin detected in indicated status of U937 Ctrl and VIMOE cells. No detected secretion can be observed. n = 3. k, THP1-derived M0 macrophages were treated with conditioned medium from THP1/U937-Ctrl/ VIMOE resting cells. The secretion level of IL-1β was detected. Two-sided unpaired t-test. n = 4. l, Experiments with Tregs directly isolated from PMBCs. Tregs were treated with conditioned medium from THP1-derived M0/M1 macrophages with anti-IL-1β neutralizing antibody/control IgG treatment. The mRNA level (left panel) and the secretion level (right panel) of IL-10 in Tregs were detected upon indicated stimulation. Two-sided unpaired t-test. n = 4 for the left panel and n = 6 for the right panel. m. Representative histogram exhibiting the CFSE signals, related to Fig. 8p. For all panels, n denotes biological replicates for each group. In e-l, data are presented as mean ± SD.
