blocking buffer  (Proteintech)

Journal: Bioactive Materials

Article Title: MSC-mimicking nanovesicle embedded bio-adhesive hydrogel for dual immunomodulation and osteogenesis to promote maxillofacial bone regeneration

doi: 10.1016/j.bioactmat.2026.02.032

Figure Lengend Snippet: nMSC@MT inheriting the original MSCs and could be effectively taken up by BMMSCs. a) Representative confocal images of PMVs with DiI-labeled cell membrane and Tht-labeled cytoplasm. Scale bar = 25 μm (above) and 1 μm (below). b) Illustration showing the co-extrusion procedure of nPMV and melatonin to obtain nMSC@MT. c) Representative TEM image of nMSC@MT. Scale bar = 100 nm. d, e) The sizes and the potential zetas of nPMV and nMSC@MT by DLS (n = 3). f, g) Coomassie brilliant blue and Western blot showing the preservation of stem cell markers, CD146, CD105, CD90, and functional protein, CXCR4, in PMV, nPMV, and nMSC@MT. h) DiI-labeled nMSC@MT (red) was detected in the cytoplasm of BMMSCs, suggesting the endocytosis of nMSC@MT. Scale bar = 10 μm. i) Immunofluorescence showing that a large amount of DiI-labeled nMSC@MT (red), released from the hydrogel, was taken up by CD146-labeled MSCs (green). Scale bar = 50 μm. j) BMMSCs uptake of DiI-labeled nMSC@MT in presence of blocking antibodies (anti-CXCR4) detected via flow cytometry. k) Analysis of the percentage of DiI positive cells in BMMSCs by flow cytometry (n = 3). l, m) Immunofluorescent staining images and corresponding semi-quantitative analysis (n = 3) of the uptake of DiI-labeled nPMV by BMMSCs in presence of blocking antibodies (anti-CXCR4). Scale bar = 50 μm. P-values are calculated using one-way ANOVA with Tukey's test, ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, n.s. not significant ( b was created with bioRender. com).

Article Snippet: To evaluate the involvement of CXCR4 in the uptake of nPMV, prior to co-culture with BMMSCs, nPMV was pre-treated with 160 nM CXCR4 blocking antibody (Ulocuplumab, Cat. HY-P99272, MedChemExpress, China) at room temperature for 1 h. Then nPMV was introduced into BMMSC cultures and incubated for 24 h. Subsequent analyses included flow cytometric analysis and immunofluorescence staining. nPMV without blocking antibody was served as a control.

Techniques: Labeling, Membrane, Western Blot, Preserving, Functional Assay, Immunofluorescence, Blocking Assay, Flow Cytometry, Staining

Journal: Genes & Diseases

Article Title: Chloride channel accessory 4 suppresses stem cell-like properties of colorectal cancer and enhances anti-PD-1 immunotherapy

doi: 10.1016/j.gendis.2025.101859

Figure Lengend Snippet: CLCA4 overexpression enhanced the therapeutic effect of anti-PD-1. (A) In vivo tumorigenicity assay in nude mice was performed to detect the therapeutic effect of CLCA4 overexpression combined with anti-PD-1 antibody. (B) Tumor growth curve was monitored from nude mice with different treatment groups. (C) Weights of tumors from nude mice with different treatments were detected. One-way ANOVA with Tukey's multiple comparisons test (mean ± standard deviation). (D) Analysis of the survival times of mice in each group ( n = 8 per group), and the experiment was terminated 60 days after tumor inoculation. Unpaired two-tailed t -test (mean ± standard deviation). (E) Immunofluorescence or immunohistochemistry staining was performed to detect the infiltration levels of CD8 + T cells, GZMB, Perforin + cells, Ki67 + , Bmi-1 + , and Oct4 + cells in tumors from different treatment groups. One-way ANOVA with Tukey's multiple comparisons test (mean ± standard deviation). (F) Hematoxylin-eosin staining of liver metastases in each group. (G) Quantitative analysis of the liver weight in each group. One-way ANOVA with Tukey's multiple comparisons test (mean ± standard deviation). (H) Quantitative analysis of the liver metastasis area/total liver area in each group. One-way ANOVA with Tukey's multiple comparisons test (mean ± standard deviation).

Article Snippet: After 7 days, mice were intraperitoneally treated with either an in vivo blocking antibody against mouse PD-1 (Clone: 29F.1A2, BioXcell, Cat# BP0273) or a rat IgG2a isotype control antibody (Clone: 2A3, BioXcell, Cat# BP0089).

Techniques: Over Expression, In Vivo, Tumorigenicity Assay, Standard Deviation, Two Tailed Test, Immunofluorescence, Immunohistochemistry, Staining