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: Cell Signaling Technology (CST) provided antibodies against cleaved caspase-1 (no. 89332), caspase-3 (no. 9661), caspase-8 (no. 4790), cleaved caspase-8 (no. 8592), cleaved GSDMD (no. 10137), ASC-Alexa Fluor 488 (no. 17507), ZBP1 (no. 33402), cGAS (no. 31659), STING (no. 13647), phospho-STING (no. 72971), TBK1 (no. 3504), phospho-TBK1 (no. 5483), phospho-IRF3 (no. 4302), IRF3 (no. 4947), and RIPK3 (no. 95702).

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

Journal: Science Advances

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

doi: 10.1126/sciadv.aee4577

Figure Lengend Snippet: ( A ) Fluorescence images of ZHDG internalization in MFC cells. ( B ) Quantification of the intracellular ZHDG fluorescence intensity. ( C ) Viability of MFC cells across treatment groups. ( D ) Annexin V–FITC/PI staining (fluorescence and flow cytometry). ( E ) TEM images of ultrastructural damage in MFC cells. ( F ) Relative LDH release. ( G ) Intracellular ATP levels. ( H ) Immunofluorescence analysis of CRT exposure and HMGB1 release. ( I ) Zinpyr-1 staining in different treatment groups. ( J ) Quantification of JC-1 aggregates and monomers. ( K ) JC-1 staining (flow cytometry). ( L ) Extracellular dsDNA release. ( M ) Western blot analysis of necroptosis markers (RIPK1, p-RIPK1, MLKL, and p-MLKL) and AIM2. ( N ) Western blot analysis of pyroptosis markers (Caspase-1, GSDMD, and cleaved forms). ( O ) Western blot analysis of apoptosis executors (Caspase-8, Caspase-3, and cleaved forms). ( P ) Western blot analysis of PANoptosis-related proteins in si AIM2 MFC cells treated with ZHDG + NIR. ( Q ) Schematic of ZHDG under NIR photoactivation inducing AIM2-PANoptosome formation and ICD. Data in (B), (C), (F), (G), and (L) are mean ± SD. Statistical analysis for (B), (C), (F), (G), and (L) was performed using a two-tailed unpaired Student’s t test. (Q) was created in BioRender. Fei, G. (2026) https://BioRender.com/wcbo7ma .

Article Snippet: Cell Signaling Technology (CST) provided antibodies against cleaved caspase-1 (no. 89332), caspase-3 (no. 9661), caspase-8 (no. 4790), cleaved caspase-8 (no. 8592), cleaved GSDMD (no. 10137), ASC-Alexa Fluor 488 (no. 17507), ZBP1 (no. 33402), cGAS (no. 31659), STING (no. 13647), phospho-STING (no. 72971), TBK1 (no. 3504), phospho-TBK1 (no. 5483), phospho-IRF3 (no. 4302), IRF3 (no. 4947), and RIPK3 (no. 95702).

Techniques: Fluorescence, Staining, Flow Cytometry, Immunofluorescence, Analysis, Western Blot, Two Tailed Test

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 DEGs in MFC cells (Control versus ZHDG + NIR). ( B ) KEGG pathway analysis. ( C ) Immunofluorescence showing ASC speck colocalization with AIM2, ZBP1, Caspase-1, Caspase-8, and RIPK1. Scale bars, 5 μm (overview) and 1 μm (enlarged panels). ( D ) Violin plot of cGAS expression across treatment groups. ( E ) Western blot analysis of cGAS-STING pathway proteins (cGAS, STING, p-STING, TBK1, p-TBK1, IRF3, and p-IRF3). ( F ) CIBERSORT analysis of immune cell infiltration. ( G ) BMDC maturation assessed by flow cytometry of CD80 + CD86 + cells. ( H ) Schematic of NIR-activated ZHDG initiating the STING-AIM2-PANoptosome cascade to potentiate antitumor immunity. Statistical analysis for (D) and (F) was performed using a two-tailed unpaired Student’s t test. (H) was created in BioRender. Fei, G. (2026) https://BioRender.com/m2dnb22 . * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cell Signaling Technology (CST) provided antibodies against cleaved caspase-1 (no. 89332), caspase-3 (no. 9661), caspase-8 (no. 4790), cleaved caspase-8 (no. 8592), cleaved GSDMD (no. 10137), ASC-Alexa Fluor 488 (no. 17507), ZBP1 (no. 33402), cGAS (no. 31659), STING (no. 13647), phospho-STING (no. 72971), TBK1 (no. 3504), phospho-TBK1 (no. 5483), phospho-IRF3 (no. 4302), IRF3 (no. 4947), and RIPK3 (no. 95702).

Techniques: Control, Analysis, Immunofluorescence, Expressing, Western Blot, Immune Cell, Flow Cytometry, Two Tailed Test