Fig. 3
S. aureus triggers mitophagy in BMDMs. (A) Heatmap displaying clustered global protein expression profiles (n = 3). (B) Volcano plot comparing differential protein expression between BMDMs infected with S. aureus for 24 h and uninfected groups. Proteins with| log2(fold change)| ≥1 and P < 0.05 are identified as significantly differentially expressed. (C) Bubble plot derived from KEGG pathway enrichment analysis. The enrichment factor indicates the degree of enrichment, while larger p-value (-log10) denote higher statistical significance. Larger bubbles signify greater enrichment levels. (D) GO enrichment analysis for proteins expressed differentially. The enrichment factor quantifies the level of enrichment. (E and F) Flow cytometry analysis of mtROS levels in BMDMs infected with S. aureus (MOI = 10) at various time points. Quantitative assessment is based on mean fluorescence intensity (MFI) indicative of intracellular mtROS levels. (G and H) Using confocal microscopy to observe changes in mitochondrial membrane potential (ΔΨm) in BMDMs infected with S. aureus (MOI = 10), at various time points after JC-1 staining. Red fluorescence indicates JC-1 aggregates, reflective of intact mitochondrial membrane potential, while green fluorescence signifies JC-1 monomers, indicating ΔΨm dissipation (Scale bar: 10 μm). Quantitative analysis of the red/green fluorescence intensity ratio reflects the overall state of mitochondrial membrane potential within the cells. (I and J) Employing JC-1 staining followed by flow cytometry to assess changes in ΔΨm. The analysis quantifies the percentage of cells positive for JC-1 monomers. (K and L) Confocal microscopy images depicting mitochondria (Mito-tracker, red), lysosomes (Lyso-tracker, green), and nuclei (Hochst, blue) in BMDMs (Scale bar: 10 μm). Histogram profiles are created by quantifying the fluorescence intensity along designated paths in the red (Mito-tracker) and green (Lyso-tracker) channels. Statistical analysis was performed using one-way ANOVA: ns, P > 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001. The data represent the mean ± SD from at least three independent experiments