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( A ) Volcano plot of differentially expressed genes (DEGs) in MFC cells (Control versus DDP + Zn 2+ ). ( B ) KEGG pathway enrichment of DEGs. ( C ) Quantification of extracellular dsDNA. ( D ) JC-1 staining assessed by fluorescence microscopy and flow cytometry. ( E <t>)</t> <t>γ-H2AX</t> immunofluorescence staining. Scale bar, 20 μm. ( F ) ASC speck colocalization with AIM2, Caspase-1, Caspase-8, and RIPK1. Scale bars, 5 μm (overview) and 1 μm (enlarged). ( G ) Western blot analysis of necroptosis markers (RIPK1, p-RIPK1, MLKL, and p-MLKL) and PANoptosis-related proteins (AIM2 and ZBP1). ( H ) Western blot analysis of apoptosis executors (Caspase-8, Caspase-3, and cleaved forms). ( I ) Western blot analysis of pyroptosis markers (Caspase-1, GSDMD, and cleaved forms). ( J ) Cell viability after DDP + Zn 2+ combined with VX-765, Ac-DEVD-CHO, or Nec-1. ( K ) PANoptosis-related proteins in si AIM2 cells. ( L and M ) cGAS activity (L) and expression (M) under Zn 2+ treatment. ( N ) Violin plot of cGAS expression. ( O ) Western blot analysis of STING pathway (STING, p-STING, TBK1, p-TBK1, IRF3, and p-IRF3). ( P ) CIBERSORT analysis of immune cell infiltration. ( Q ) BMDC maturation (CD80 + CD86 + ) after different treatments. ( R ) Schematic of the DDP + Zn 2+ –induced ICD. Data in (C), (J), and (L) are mean ± SD. Statistical analysis for (C), (J), (L), (N), and (P) used a two-tailed unpaired Student’s t test. (R) was created in BioRender. Fei, G. (2026) https://BioRender.com/8ri399r . * P < 0.05.
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( A ) Volcano plot of differentially expressed genes (DEGs) in MFC cells (Control versus DDP + Zn 2+ ). ( B ) KEGG pathway enrichment of DEGs. ( C ) Quantification of extracellular dsDNA. ( D ) JC-1 staining assessed by fluorescence microscopy and flow cytometry. ( E <t>)</t> <t>γ-H2AX</t> immunofluorescence staining. Scale bar, 20 μm. ( F ) ASC speck colocalization with AIM2, Caspase-1, Caspase-8, and RIPK1. Scale bars, 5 μm (overview) and 1 μm (enlarged). ( G ) Western blot analysis of necroptosis markers (RIPK1, p-RIPK1, MLKL, and p-MLKL) and PANoptosis-related proteins (AIM2 and ZBP1). ( H ) Western blot analysis of apoptosis executors (Caspase-8, Caspase-3, and cleaved forms). ( I ) Western blot analysis of pyroptosis markers (Caspase-1, GSDMD, and cleaved forms). ( J ) Cell viability after DDP + Zn 2+ combined with VX-765, Ac-DEVD-CHO, or Nec-1. ( K ) PANoptosis-related proteins in si AIM2 cells. ( L and M ) cGAS activity (L) and expression (M) under Zn 2+ treatment. ( N ) Violin plot of cGAS expression. ( O ) Western blot analysis of STING pathway (STING, p-STING, TBK1, p-TBK1, IRF3, and p-IRF3). ( P ) CIBERSORT analysis of immune cell infiltration. ( Q ) BMDC maturation (CD80 + CD86 + ) after different treatments. ( R ) Schematic of the DDP + Zn 2+ –induced ICD. Data in (C), (J), and (L) are mean ± SD. Statistical analysis for (C), (J), (L), (N), and (P) used a two-tailed unpaired Student’s t test. (R) was created in BioRender. Fei, G. (2026) https://BioRender.com/8ri399r . * P < 0.05.
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( A ) Volcano plot of differentially expressed genes (DEGs) in MFC cells (Control versus DDP + Zn 2+ ). ( B ) KEGG pathway enrichment of DEGs. ( C ) Quantification of extracellular dsDNA. ( D ) JC-1 staining assessed by fluorescence microscopy and flow cytometry. ( E ) γ-H2AX immunofluorescence staining. Scale bar, 20 μm. ( F ) ASC speck colocalization with AIM2, Caspase-1, Caspase-8, and RIPK1. Scale bars, 5 μm (overview) and 1 μm (enlarged). ( G ) Western blot analysis of necroptosis markers (RIPK1, p-RIPK1, MLKL, and p-MLKL) and PANoptosis-related proteins (AIM2 and ZBP1). ( H ) Western blot analysis of apoptosis executors (Caspase-8, Caspase-3, and cleaved forms). ( I ) Western blot analysis of pyroptosis markers (Caspase-1, GSDMD, and cleaved forms). ( J ) Cell viability after DDP + Zn 2+ combined with VX-765, Ac-DEVD-CHO, or Nec-1. ( K ) PANoptosis-related proteins in si AIM2 cells. ( L and M ) cGAS activity (L) and expression (M) under Zn 2+ treatment. ( N ) Violin plot of cGAS expression. ( O ) Western blot analysis of STING pathway (STING, p-STING, TBK1, p-TBK1, IRF3, and p-IRF3). ( P ) CIBERSORT analysis of immune cell infiltration. ( Q ) BMDC maturation (CD80 + CD86 + ) after different treatments. ( R ) Schematic of the DDP + Zn 2+ –induced ICD. Data in (C), (J), and (L) are mean ± SD. Statistical analysis for (C), (J), (L), (N), and (P) used a two-tailed unpaired Student’s t test. (R) was created in BioRender. Fei, G. (2026) https://BioRender.com/8ri399r . * P < 0.05.

Journal: Science Advances

Article Title: Chemo-metallo nanodrivers for spatiotemporally controlled in situ vaccination

doi: 10.1126/sciadv.aee4577

Figure Lengend Snippet: ( A ) Volcano plot of differentially expressed genes (DEGs) in MFC cells (Control versus DDP + Zn 2+ ). ( B ) KEGG pathway enrichment of DEGs. ( C ) Quantification of extracellular dsDNA. ( D ) JC-1 staining assessed by fluorescence microscopy and flow cytometry. ( E ) γ-H2AX immunofluorescence staining. Scale bar, 20 μm. ( F ) ASC speck colocalization with AIM2, Caspase-1, Caspase-8, and RIPK1. Scale bars, 5 μm (overview) and 1 μm (enlarged). ( G ) Western blot analysis of necroptosis markers (RIPK1, p-RIPK1, MLKL, and p-MLKL) and PANoptosis-related proteins (AIM2 and ZBP1). ( H ) Western blot analysis of apoptosis executors (Caspase-8, Caspase-3, and cleaved forms). ( I ) Western blot analysis of pyroptosis markers (Caspase-1, GSDMD, and cleaved forms). ( J ) Cell viability after DDP + Zn 2+ combined with VX-765, Ac-DEVD-CHO, or Nec-1. ( K ) PANoptosis-related proteins in si AIM2 cells. ( L and M ) cGAS activity (L) and expression (M) under Zn 2+ treatment. ( N ) Violin plot of cGAS expression. ( O ) Western blot analysis of STING pathway (STING, p-STING, TBK1, p-TBK1, IRF3, and p-IRF3). ( P ) CIBERSORT analysis of immune cell infiltration. ( Q ) BMDC maturation (CD80 + CD86 + ) after different treatments. ( R ) Schematic of the DDP + Zn 2+ –induced ICD. Data in (C), (J), and (L) are mean ± SD. Statistical analysis for (C), (J), (L), (N), and (P) used a two-tailed unpaired Student’s t test. (R) was created in BioRender. Fei, G. (2026) https://BioRender.com/8ri399r . * P < 0.05.

Article Snippet: DNA damage was assessed using a γ-H2AX Immunofluorescence Assay Kit, Fluo-4 AM, and Hoechst (Beyotime Biotechnology, Shanghai, China).

Techniques: Control, Staining, Fluorescence, Microscopy, Flow Cytometry, Immunofluorescence, Western Blot, Analysis, Activity Assay, Expressing, Immune Cell, Two Tailed Test