va7 2 3c10 standard biotools 3153024b 159tb cd197  (fluidigm)

Journal: Science Advances

Article Title: Nanocollision promotes locomotion of dendritic cells for tumor therapy

doi: 10.1126/sciadv.aeb7714

Figure Lengend Snippet: ( A ) Synthesis process of an NCG. (PFH, liquid-gas phase-change core; PDA, stabilizer; FA, solid-liquid phase-change shell) capable of gas-liquid-solid triphasic conversion. ( B ) Schematic of the strategy for NCG-triggered nanocollision for enhanced locomotion of DCs. First, ultrasound induces vaporization of PFH. Then, the gasified PFH generates a critical internal pressure, triggering instantaneous rupture of the FA shell with subsequent outward ejection of fragmented particulates. Subsequently, the fragmented particulates induce nanocollisions with DCs, eliciting localized fluctuation of plasma membrane. IV, Piezo1 detects the fluctuation and mediates Ca 2+ influx through its central pore. Finally, Ca 2+ influx induces F-actin polymerization (enhanced intrinsic locomotion) and high expression of CCR7 (enhanced chemotaxis). ( C ) Locomotion-enhanced DCs potentiate antigen capture and lymph node homing, thereby activating T cells to amplify antitumor immunity.

Article Snippet: Antibodies and reagents used for Western blot are as follows: rabbit anti–IL-12 (bs-0767R), rabbit anti–IFN-γ (bs-0480R), CCR7 rabbit polyclonal antibody (pAb) (bs-1305R), and β-actin mouse monoclonal antibody (bsm-33036 M) from Bioss; cPLA2 pAb (YT1084), cPLA2 (phospho Ser 505 ) pAb (YP0868), PIEZ1 rabbit pAb (YT8073), AP-1 (phospho Tyr 170 ) pAb (YP0018), and AP-1 (Acetyl Lys271) pAb (YK0062) from Immunoway; anti-calreticulin rabbit pAb ( GB112134 ), anti-HMGB1 rabbit pAb (GB11103) from Servicebio.

Techniques: Clinical Proteomics, Membrane, Expressing, Chemotaxis Assay

Journal: Science Advances

Article Title: Nanocollision promotes locomotion of dendritic cells for tumor therapy

doi: 10.1126/sciadv.aeb7714

Figure Lengend Snippet: ( A and B ) Representative flow cytometry dot plots (A) and percentage (B) of CFSE-stained DCs in adjacent lymph nodes. ( n = 5). ( C ) Western blot analysis of CCR7 expression and oligomerization. ( D ) cPLA 2 pathway–related gene alterations, heatmap. (C versus B). ( E and F ) Western blot analysis of expression for proteins in the cPLA 2 pathway, including Piezo1, p-cPLA 2 , CCR7, and their quantitative analysis. ( G ) Representative images of Ca 2+ diffusion from the protrusions to the cell interior. ( H ) Schematic illustration of Ca 2+ influx–induced CCR7 expression in DC. ( I ) Heatmap of collagen and integrin-related genes (A versus B versus C). ( J ) Sankey bubble plot of pathway changes corresponding to genes. (C versus B). ( K ) Density plot of RNA-seq. (A versus B versus C). ( L ) Bubble plot of nanocollision-mediated alterations in DC signaling cascades. (C versus B). ( M and N ) Representative fluorescence images (M) and statistical graphs (N) of deep infiltration of DCs. (A, control; B, magnetic nanospheres; C, magnetic nanospheres with magnetic field, which can achieve collision. n = 6) ( P values: ns, not significant, * P < 0.05, ** P < 0.01.)

Article Snippet: Antibodies and reagents used for Western blot are as follows: rabbit anti–IL-12 (bs-0767R), rabbit anti–IFN-γ (bs-0480R), CCR7 rabbit polyclonal antibody (pAb) (bs-1305R), and β-actin mouse monoclonal antibody (bsm-33036 M) from Bioss; cPLA2 pAb (YT1084), cPLA2 (phospho Ser 505 ) pAb (YP0868), PIEZ1 rabbit pAb (YT8073), AP-1 (phospho Tyr 170 ) pAb (YP0018), and AP-1 (Acetyl Lys271) pAb (YK0062) from Immunoway; anti-calreticulin rabbit pAb ( GB112134 ), anti-HMGB1 rabbit pAb (GB11103) from Servicebio.

Techniques: Flow Cytometry, Staining, Western Blot, Expressing, Diffusion-based Assay, RNA Sequencing, Fluorescence, Control

Journal: Science Advances

Article Title: Nanocollision promotes locomotion of dendritic cells for tumor therapy

doi: 10.1126/sciadv.aeb7714

Figure Lengend Snippet: ( A and B ) Representative fluorescence images of intracellular Ca 2+ distribution in DCs with different treatments. ( C to E ) Locomotion trajectory (C), accumulated distance (in 5 min) (D), and locomotion speed (E) of DCs with different treatments. ( F and G ) Western blot analysis of the nanocollision-induced alterations of monomeric and oligomeric CCR7 (F) and Piezo1 expression (G). ( H and I ), Representative fluorescence images showing the antigen capture (H) and antigen presenting (I) capabilities of DCs with different treatments. ( J ) Western blot analysis of IL-12 and IFN-γ expression. ( K to N ) Transcriptomics analysis of gene expression in DCs. RNA-seq density plot [(K), A versus B versus C], heatmap of immune activation-related genes [(L) A versus B versus C], bubble plot [(M) C versus B], and sankey bubble plot [(N) C versus B]. [(A) Control; (B) R848; (C) R848 + NCG US(40°C) ]. P values: ns, not significant, * P < 0.05, ** P < 0.01, **** P < 0.0001.)

Article Snippet: Antibodies and reagents used for Western blot are as follows: rabbit anti–IL-12 (bs-0767R), rabbit anti–IFN-γ (bs-0480R), CCR7 rabbit polyclonal antibody (pAb) (bs-1305R), and β-actin mouse monoclonal antibody (bsm-33036 M) from Bioss; cPLA2 pAb (YT1084), cPLA2 (phospho Ser 505 ) pAb (YP0868), PIEZ1 rabbit pAb (YT8073), AP-1 (phospho Tyr 170 ) pAb (YP0018), and AP-1 (Acetyl Lys271) pAb (YK0062) from Immunoway; anti-calreticulin rabbit pAb ( GB112134 ), anti-HMGB1 rabbit pAb (GB11103) from Servicebio.

Techniques: Fluorescence, Western Blot, Expressing, Gene Expression, RNA Sequencing, Activation Assay, Control