Fig. 1: Lysosomal protease leakage triggers mitochondrial protein degradation in macrophages.

a Schematic showing the conditions tested in this study. b Immunoblots for MFN2, TOM20, TIM23, HSP60 and Citrate synthase (CS) in iPSDM stimulated with 0.5 mM LLOMe for 1 h, 100 μg/mL silica crystals or beads for 3 h or infected with Mtb WT or Mtb ΔRD1 for 48 h and incubated with the indicated protease or proteasome inhibitors. Beta-actin (ACTB) levels were used as loading controls (repeated three times with similar results). c Immunoblots for mitochondrial proteins in iPSDM WT, ATG7 KO, PRKN KO and PRKN/ATG7 DKO stimulated with 0.5 mM LLOMe for 1 h and incubated in the presence or absence of the indicated inhibitors (repeated three times with similar results). d Immunoblots for mitochondrial proteins in BMM WT, CtsB KO, CtsL KO and CtsS KO stimulated with 0.5 mM LLOMe for 1 h. (repeated three times with similar results). e Representative images of iPSDM expressing the mitophagy reporter NIPSNAP and stimulated with 0.5 mM LLOMe for 1 h, 100 μg/mL silica crystals for 3 h, infected with Mtb WT for 48 h or treated with 20 μM CCCP for 3 h. f NIPSNAP mCherry only puncta evaluated by confocal microscopy, n = 30 cells examined per condition over three independent experiments. g TOM20⁺/PDHA1⁻ and PDH⁺/TOM20⁻ MDVs were quantified after the indicated conditions. Glucose oxidase (GO) was used at 50 mU/ml for 1 h as a positive control, n = 20 cells examined per condition over three independent experiments. h One‐way ANOVA and Tukey post-test was used for multiple comparisons ***p ≤ 0.001, ns no significant. Scale bars, 10 μm. Floating bar plots show minimum and maximum values, line at mean. Unprocessed blots and Source data are provided as a Source Data file.