Fig. 5: ZEB1 supports inflammatory macrophage polarization and cytokine secretion by regulating protein biosynthesis, trafficking and cytokine release.
From: Macrophages foster anti-tumor immunity by ZEB1-dependent cytotoxic T cell chemoattraction
Venn diagrams of differentially expressed genes (DEGs) of LysMCtrl (blue) and LysMΔZeb1 BMDMs (red) after stimulation with LPS (a) or IL-4 (b) compared to unstimulated in total (left) and divided in up-/downregulated DEGs (right). c GO term enrichment analysis for DEGs (FDR < 0.05) uniquely up- or downregulated by LysMΔZeb1 BMDMs after LPS stimulation. d Log2 fold change of expression of selected trafficking genes after LPS stimulation. X marks no significant deregulation. e Representative images and quantification of OPP incorporation of LysMCtrl and LysMΔZeb1 BMDMs with 0 h, 4 h and 16 h LPS pre-stimulation (n = 3; means ± SD). f Transmission electron microscopy of LPS-stimulated LysMCtrl and LysMΔZeb1 BMDMs with representative images (top) and scoring of indicated vesicle types (bottom) from 26 (LysMCtrl) and 25 (LysMΔZeb1) cells derived from 3 independent BMDM lines per genotype. (means ± SD;). g Quantification of intracellular cytokine levels in pg/ml per 10 µg of protein lysates obtained from untreated (M0) or LPS-treated LysMCtrl and LysMΔZeb1 BMDMs using a bead-based immunoassay (n = 3; means ± SD). h–i Western blots of the indicated trafficking factors and β-Actin as loading control of protein lysates from independent LysMCtrl and LysMΔZeb1 BMDM lines either unstimulated (M0) or stimulated with LPS (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 ns: not significant; 2-way ANOVA (e, g), Mann-Whitney (f)). Note that in (e), color-coded indicators (blue/red = LysMCtrl/LysMΔZEB1) on top of the bars indicate comparisons of a given time point type to the respective time point “0 h” within one genotype and comparisons between LysMCtrl and LysMΔZEB1 at specific time points are marked by the connectors. j Hypothetical model for ZEB1-related cargo trafficking in BMDMs leading to reduced cytokine release (MVBs: Multivesicular bodies). Thickness of the arrows indicates the anticipated trafficking strength. Created in BioRender. Schuhwerk, H. (2025) https://BioRender.com/eymhtmg.
