Fig. 2: ZEB1 regulates mitochondrial content and autophagy during inflammation.

a MTG staining in peritoneal macrophages from wild-type mice untreated, treated with LPS, or treated with LPS + LPS. The FACS plot is representative of at least 3 independent experiments. b MTG staining in the monocyte-enriched PBMC of healthy controls and septic patients at 0 h and 72 h. The FACS plot is representative of at least 5 independent experiments. c Western blot for MT-CO1 and GAPDH in PBMCs from three healthy donors and three septic patients at 0 and 72 h. The blot is representative of three independent experiments. d MT-CO1 protein levels in five healthy controls and five patients relative to GAPDH. e TEM of mitochondria in sorted macrophages from Zeb1^WT (labeled in blue) and Zeb1^∆M (orange) mice treated with PBS (Untreated), LPS for the indicated periods, or LPS + LPS. A representative macrophage from a 4–5 mice pool for each genotype and condition at 15,000X and 40,000X magnification. Scale bar: 2 μm. f At least 6 pictures were quantified for each genotype and condition. (n = 7,6,6,8,9,10,6,8,10,7) g Heatmap of autophagy-related genes in peritoneal macrophages from Zeb1^WT and Zeb1^∆M mice treated with LPS or LPS + LPS. h Sqstm1 mRNA in peritoneal macrophages from Zeb1^WT and Zeb1^∆M untreated, treated with LPS or LPS + LPS. (n = 5,4,6,4,4,4). i As in Fig. 2c, but for p62/SQSTM1 and β-ACTIN. Blots are representative of four independent experiments. j Ultrastructure of autophagic vacuoles in LPS-treated macrophages. TEM images of Zeb1^WT and Zeb1^∆M macrophages from mice treated with LPS for 3 h. Yellow asterisks: autophagic vacuoles in Zeb1^WT macrophages, two with increased electron density (autolysosomes). N nucleus, M mitochondria, ER endoplasmic reticulum. Scale bar: 1000 nm. A representative macrophage from a 4–5 mice pool for each genotype and condition. At least 6 pictures were analyzed for each genotype and condition. Graph bars in Fig.  2 represent mean values +/− SEM with two-tailed unpaired Mann–Whitney test. p ≤ 0.001 (***), p ≤ 0.01 (**) or p ≤ 0.05 (*) levels, or non-significant (ns) for values of p > 0.05. Raw data along p values for statistical analyses are included in the Source Data file.