Journal: Science Advances

Article Title: Cold atmospheric plasma–engineered nanovaccine with spatiotemporal sequential immunization reprograms antitumor immunity

doi: 10.1126/sciadv.aeb5894

Figure Lengend Snippet: ( A ) The preparation of CAPTURE. ( B and C ) Representative TEM image and dynamic light scattering (DLS) showed NVs of similar shapes and sizes. Scale bars, 200 nm. ( D to I ) OVA-specific expansion and activation of primary mouse T cells: 10 5 OT-I CD8 + T cells per well were coincubated with a variety of NVs (20 μg ml −1 ) (D), including CAPTURE with different constructs of the αCD28 antibody (E) or different type of vesicles (F to I). Following 3 days of coincubation, assessments of T cell expansion (F) and activation (G to I) were conducted. ( J ) Cytotoxicity of OT-I CD8 + T cells against B16-OVA cells post–NVs treatment under various conditions; cells were coincubated at differing effector-target (E:T) ratios for 24 hours. ( K ) After 36 hours of CAP treatment, the induced expression of integrin α6, integrin β4, and v-SNARE in tumor cell NVs was evaluated by Western blot analysis. ( L ) Confocal microscopy images of B16-OVA tumor cells incubated with CAPTURE (20 μg ml −1 ) for 24 hours. Blue: cell nuclei, green: MHC-I, red: Flag tag. Scale bar, 10 μm. (B), (C), (K), and (L) show representative results of two independent experiments with similar results. The data are shown as the mean ± SD from a representative experiment of two to three independent experiments with n = 5 (E, F, and I) and n = 4 (G, H, and J) biologically independent samples. Statistical significance was calculated via one-way ANOVA in (E) and (G) to (I) or two-way ANOVA in (J) with Bonferroni multiple comparisons posttest. Synergy analysis (via the Highest Single Agent model) comparing the B16-OVA-NV(CAP), B16-OVA-NV-αCD28, and CAPTURE groups indicated a significant synergistic effect for CAPTURE [ P = 0.0102 (G), P = 0.0015 (H), and P = 0.0026 (I)].

Article Snippet: The expression of Integrin α6, Integrin β4, and v-SNARE (all antibodies sourced from Santa Cruz Biotechnology) was analyzed by Western blot.

Techniques: Activation Assay, Construct, Expressing, Western Blot, Confocal Microscopy, Incubation, FLAG-tag

Journal: The Journal of Biological Chemistry

Article Title: A disintegrin and metalloproteinase with thrombospondin motifs 18 (ADAMTS18) cleaves fibronectin and negatively regulates its fibrillogenesis

doi: 10.1016/j.jbc.2025.110844

Figure Lengend Snippet: Effect of ADAMTS18 knockdown on gene expression. A , heatmap representing significant DEGs ( p adj <0.05) common to siAD18-1, siAD18-2, and siAD18-3 at 24 h ( left ) and 96 h ( right ) after transfection. B , Gene Ontology (GO) analysis (g:Profiler) of significant DEGs common to siAD18-1, siAD18-2, and siAD18-3 either at 24 h or 96 h after transfection. C , RT–quantitative PCR analysis of LAMA5 and RBP J expression 24 h after siCTRL, siAD18-1, siAD18-2, and siAD18-3 transfection in HUVEC (mean ± SD, n = 4, lognormal repeated-measures one-way ANOVA with Tukey’s multiple comparison test). D , RT–quantitative PCR analysis of CXCL1 and ITGB4 expression 96 h after siCTRL, siAD18-1, siAD18-2, and siAD18-3 transfection in HUVEC (mean ± SD, n = 4, lognormal repeated-measures one-way ANOVA with Tukey’s multiple comparison test). E , Western blot (WB) analysis of siCTRL-, siAD18-1-, siAD18-2-, and siAD18-3-transfected HUVEC with anti-ITGB4 antibody (1:250 dilution, sc-514426; Santa Cruz) 96 h after transfection. The siCTRL lane was reordered to the left side of the blot from the right side. F , densitometry analysis (Image Lab) of ITGB4 205 kDa and GAPDH 36 kDa band intensities. The adjusted volume (Int) for ITGB4 was normalized to an adjusted volume (Int) of GAPDH. The data are presented as fold change of siAD18 to siCTRL (mean ± SD, n = 3 using two different HUVEC donors, ordinary one-way ANOVA). Significance is indicated as follows: ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001. ADAMTS18, A disintegrin and metalloproteinase with thrombospondin motifs 18; DEG, differentially expressed gene; HUVEC, human umbilical vein endothelial cell.

Article Snippet: E , Western blot (WB) analysis of siCTRL-, siAD18-1-, siAD18-2-, and siAD18-3-transfected HUVEC with anti-ITGB4 antibody (1:250 dilution, sc-514426; Santa Cruz) 96 h after transfection.

Techniques: Knockdown, Gene Expression, Transfection, Real-time Polymerase Chain Reaction, Expressing, Comparison, Western Blot