Journal: bioRxiv

Article Title: Development of Integrin α5β1-targeted PET/NIR imaging probes for glioblastoma intraoperative navigation and intracavity targeted radionuclide therapy

doi: 10.64898/2026.01.09.698741

Figure Lengend Snippet: (A) Strategies for modification of GS, including amino acid mutation, covalent modification, cyclization, and multivalency. (B) SPR sensorgrams demonstrating the binding affinity of GS and GR for human integrin α5β1 in a concentration-dependent manner. The equilibrium dissociation constant (K D ) of each peptide was calculated based on SPR measurements. The K D values of each precursor are shown. (C) Molecular docking of GR and GS binding integrin a5b1 protein (grey; PDB: 7NWL) showing the selected possible ligation residues. (D-F) Analysis of integrin α5β1 expression in U87MG cells and tumor tissues by western blot ( D ), and immunohistochemistry ( E-F ) analysis. The band for integrin α5 was approximately 150 kDa. M, marker. C, cell. T, tumor. For immunofluorescence images, green is for integrin α5, red for integrin β1, and blue for nucleus. Scale bar, 50□μm ( E-F ). (G) In vitro cellular uptake of [ 68 Ga]GS and [ 68 Ga]GR in U87MG cell lines. All results are expressed as means ± SEM, as indicated in at least three independent experiments. “*” represents differences compared with the [ 68 Ga]GS. * p < 0.05.

Article Snippet: Recombinant human integrin α5β1 (alpha 5 beta 1, HY-P77718, MCE, NJ, USA) was immobilized on a CM5 sensor chip at 25 □ following a standard amine coupling kit.

Techniques: Modification, Mutagenesis, Binding Assay, Concentration Assay, Ligation, Expressing, Western Blot, Immunohistochemistry, Marker, Immunofluorescence, In Vitro

Journal: bioRxiv

Article Title: Development of Integrin α5β1-targeted PET/NIR imaging probes for glioblastoma intraoperative navigation and intracavity targeted radionuclide therapy

doi: 10.64898/2026.01.09.698741

Figure Lengend Snippet: (A) Immunofluorescence images show the expression of integrin α5β1, GFAP, and NEUN of U87MG tumors in orthotopic glioblastoma tumor-bearing mice. Green indicates integrin α5, Red for NEUN, yellow for GFAP, and blue for nucleus. Scale bar, 1 mm. (B-C) Representative fluorescence imaging of mice bearing in orthotopic U87MG tumors ( B ) and ex vivo brains ( C ). Cy5-GS and Cy5-GR were intravenously injected with a dose of 5 mg/kg. (D) Quantification of fluorescence intensity in tumors corresponding to ( C ). (E) Immunohistochemical analysis of integrin α5β1 expression in brain tissue sections from orthotopic glioblastoma tumor-bearing mice. Scale bar, 1 mm. (F-G) Immunofluorescence images of brain tissue sections from orthotopic glioblastoma tumor-bearing mice treated with Cy5-GS ( F ) and Cy5-GR ( G ). Green indicates integrin α5, Red for GS or GR, and blue for nucleus. Scale bar, 1 mm. (H-I) Magnific imaging of brain tissue sections from orthotopic glioblastoma tumor-bearing mice treated with GS-Cy5. Scale bar, 50 μm. All results are expressed as means ± SEM, as indicated in at least three independent experiments. A multiple t-test was used when two groups were compared. The symbol “*” represents differences compared with the Cy5-GS. *** p < 0.001.

Article Snippet: Recombinant human integrin α5β1 (alpha 5 beta 1, HY-P77718, MCE, NJ, USA) was immobilized on a CM5 sensor chip at 25 □ following a standard amine coupling kit.

Techniques: Immunofluorescence, Expressing, Fluorescence, Imaging, Ex Vivo, Injection, Immunohistochemical staining

Journal: Bioactive Materials

Article Title: A large puncture closer of aortic wall by multi-memory actions with thrombo-hemodynamic control

doi: 10.1016/j.bioactmat.2025.12.042

Figure Lengend Snippet: 18 Fr puncture of hemostasis in porcine aorta using VWP by validating the memory programming effect of each part. a, As a challenging model for application of large-diameter catheters, i) an 18 Fr (6 mm) puncture is created into the porcine thoracic aorta (diameter: 10 mm) so that the size-matched VWP is deployed, followed by measuring proximal and distal blood pressure. ii) The experimental groups are designed first to exam the memory programming effect of collaboration between Ring squeezing with Body expansion on self-locking (SL) to enable efficient hemostasis. Next, the effect of Wing shape recovery from curve to flat is examined on hemodynamic control (HC) in cooperation with the actions of Body and Ring to handle hemostasis. No recovery of Wing shape is expected to induce excessive thrombosis. iii) Four experimental groups are established using a total of 12 pigs (N = 12) with immediate sacrifice following deployment (N = 3 each). Group 1 [SL(−) HC(−)] represents no memory programming. Group 2 [SL(+) w/flat Wing] has the effects of Body and Ring actions except the hemostatic sealing by keeping the flat Wing. Group 3 [SL(+) HC(+)] possesses the complete memory effects of the three parts. Group 4 [SL(+) w/bump Wing] is expected to have excessive thrombosis because of no shape recovery from the curved Wing while maintaining the memory actions of Body and Wing. b, Each group is visually explained in the illustrations. c , In VWP actions, (left) the bleeding condition preserves the normal sinusoidal waveform of high proximal pressure (green) in contrast to the disturbed waveform of low distal pressure (red). (middle) Hemostatic closure results in similar high sinusoidal waveform at both pressure sites. (right) Excessive thrombosis does not disturb the waveform, but the distal pressure level becomes lower than the proximal one. d, When reperfusion starts by removing the clamp post-deployment (blue), only Group 3 [SL(+) HC(+)] reaches the hemostatic closure, as evidenced by flow stabilization (red) with a 5 s plateau at both pressure sites. Group 4 [SL(+) w/bump Wing] exhibits the pattern of over-thrombosis. e, H&E images show bleeding in Group 1 as an indication of incomplete closure in contrast to moderate, minimal, and dense thrombotic features observed in Group 2, 3, and 4 respectively as further supported by the signals of activated platelets (green, CD41-positive) and fibrinogen (red) [Scale bars = 0.5 mm (4 mm in box)]. f, Compared to Group 1 [SL(−) HC(−)] and 2 [SL(+) w/flat Wing], Group 3 [SL(+) HC(+)] shows the fastest i) hemostasis and ii) arterial pressure equilibration, indicating the most efficient hemostatic response. g, These outcomes in Group 3 include i) the smallest difference between the proximal and distal pressures with ii) the smallest thrombus area in contrast the largest area of Group 4 [SL(+) w/bump Wing] as an indication of excessive thrombosis. h , The marker gene expression of thrombotic feature (vWF, PF-4, and P-sel) significantly increases from Group 2 to Group 3 and further to Group 4 except the comparison of vWF expression between Group 2 and 3 (ns: no significance). Data are shown as mean ± SD, N = 3 biologically independent animals per group. Significance was determined using one-way ANOVA with Tukey's test between groups.

Article Snippet: Primary antibodies against CD41 (1:100, 24552-1-AP, proteintech), fibrinogen (1:100, ab232793, Abcam), CD31 (1:100, sc-376764, Santa Cruz Biotechnology), CD68 (1:100, ab125212, Abcam), and ARG-1 (1:200, LS-C447907, LSBio) were applied overnight at 4°C.

Techniques: Control, Marker, Gene Expression, Comparison, Expressing

Journal: bioRxiv

Article Title: Molecular Imaging of the TGF-β Activating Integrin αvβ6 Detects Chronic Lung Allograft Dysfunction

doi: 10.64898/2026.01.07.698265

Figure Lengend Snippet: (A) Flow cytometric analysis of POD 16-tolerant and CLAD allografts with (A) representative contour plots gated on live CD45-allograft cells (N=5/group) and (B) Intracellular Sftpc expression analysis on CLAD allograft CD326+ αvβ6+ cells. The histogram shown is representative of three independent experiments. (C) Plot of POD 16 allograft numbers of live CD45-CD326+ αvβ6+ cells shown with a mean ± standard deviation from the mean for N=5 per group. Representative immunohistochemical stains with αvβ6 specific antibodies with indicated magnification and scales (N≥4/group).

Article Snippet: Sections were then stained with 1:200 αvβ6 antibodies (bs-5791R, Bioss for mouse tissue; AB00891-23.0, Absolute Antibody for human tissue) overnight at 4°C.

Techniques: Expressing, Standard Deviation, Immunohistochemical staining

Journal: bioRxiv

Article Title: Molecular Imaging of the TGF-β Activating Integrin αvβ6 Detects Chronic Lung Allograft Dysfunction

doi: 10.64898/2026.01.07.698265

Figure Lengend Snippet: αvβ6 immunohistological staining of allografts from recipients that underwent club cell injury and received (A) CCL2 neutralizing Abs, (B) CD8α depleting Abs, respectively, and (A, B) Control Ig injections followed by POD 16 [ 64 Cu]Cu-DOTA-A20-K16R PET/CT imaging. Images shown are representative of N≥4 per group. Scale bars: 400 μm for 10x, 100 μm for 40x.

Article Snippet: Sections were then stained with 1:200 αvβ6 antibodies (bs-5791R, Bioss for mouse tissue; AB00891-23.0, Absolute Antibody for human tissue) overnight at 4°C.

Techniques: Staining, Control, Positron Emission Tomography-Computed Tomography, Imaging

Journal: bioRxiv

Article Title: Molecular Imaging of the TGF-β Activating Integrin αvβ6 Detects Chronic Lung Allograft Dysfunction

doi: 10.64898/2026.01.07.698265

Figure Lengend Snippet: (A, Left Panel) Representative autoradiographic images following incubation of [ 64 Cu]Cu-DOTA-A20-K16R with human explanted lung tissue sections from a patient with CLAD and Control lung tissue from a donor lung obtained prior to transplantation. Preincubation with an excess of a cold probe sharply reduces autoradiographic activity (Cold Probe/CLAD). (A, Right Panel) corresponding trichrome-stained lung sections that were incubated with radiotracer. Images are representative of independently conducted experiments, with (B) violin plots showing combined data from 3 explanted CLAD transplants with or without cold-probe blockade and 3 control lungs per group. Two-sided Mann-Whitney U t-test *p<0.05, ***p<0.001. Representative αvβ6 (C) immunohistochemical and (D) immunofluorescent staining of human explanted lungs with CLAD and control lungs (N=3/group). Arrows denote clusters of cells expressing αvβ6. Scale bars: 400 μm for 10x, 100 μm for 40x.

Article Snippet: Sections were then stained with 1:200 αvβ6 antibodies (bs-5791R, Bioss for mouse tissue; AB00891-23.0, Absolute Antibody for human tissue) overnight at 4°C.

Techniques: Incubation, Control, Transplantation Assay, Activity Assay, Staining, MANN-WHITNEY, Immunohistochemical staining, Expressing

Journal: eLife

Article Title: Vascular endothelial-specific loss of TGF-beta signaling as a model for choroidal neovascularization and central nervous system vascular inflammation

doi: 10.7554/eLife.107018

Figure Lengend Snippet: Retina flatmounts were stained for HIF1-alpha to visualize regions with hypoxia. Territories with accumulation of HIF1-alpha, localized to nuclei, are seen in Cdh5CreER;Tgfbr1 CKO/- retinas. Control retinas have lower levels of HIF1-alpha. Scale bar, 100 µm.

Article Snippet: The following antibodies were used for tissue immunohistochemistry: rat mAb anti-mouse PLVAP/MECA-32 (BD Biosciences 553849); rat mAb anti-mouse CD31 (BD Biosciences 553370); rat anti-mouse ICAM-1 (Invitrogen 14-0542-82); rat mAb anti-mouse F4/80 (Bio-Rad MCA497G); rat mAb anti-mouse CD206 (Bio-Rad MCA2235); rat mAb anti-mouse PU.1/Spi-1 (R&D Systems MAB7124); mouse mAb anti-alpha SMA, Cy3 conjugate (Sigma-Aldrich C6198); mouse mAb anti-CLDN5, Alexa Fluor 488 conjugate (Thermo Fisher Scientific 352588); mouse mAb anti-RPE65, Dylight 550 conjugate (Invitrogen MA5-16044); rabbit polyclonal anti-Collagen IV (Novus Biologicals NB120-6586); rabbit polyclonal anti-NG2 Chondroitin Sulfate Proteoglycan (Millipore AB5320); rabbit mAb anti-ASC/TMS1 (Cell Signaling 67824S); rabbit mAb anti-cleaved Caspase-3 (Cell Signaling 9664S); rabbit mAb anti-HIF-1alpha (Cell Signaling 36169S); rabbit mAb anti-P-SMAD1/5/9 (Cell Signaling 13820S); Armenian hamster mAb anti-CD3e (Invitrogen 14-0031-82); goat polyclonal anti-mouse CD45 (R&D Systems AF114); goat polyclonal anti-Iba1 (Novus Biologicals NB100-1028); chicken polyclonal anti-GFP (Aves Labs GFP-1020); rabbit mAb anti-NFkappaB NF-κB p65 (D14E12; Cell Signaling Technology 8242S); rabbit mAb anti-Integrin alpha 2 (ITGA2; clone GEB, BosterBio M01933); rabbit mAb anti-Integrin alpha 4 (ITGA4; D2E1; Cell Signaling Technology 8440); rabbit mAb anti-TOX/TOX2 (E6G5O; Cell Signaling Technology 36778S).

Techniques: Staining, Control