Journal: Clinical and Applied Thrombosis/Hemostasis

Article Title: Transcriptome-wide m6A Methylation Profiling Reveals Regulatory Networks During Endothelial Differentiation of Bone Marrow Mesenchymal Stem Cells

doi: 10.1177/10760296261472670

Figure Lengend Snippet: Isolation and characterization of human bone marrow-derived mesenchymal stem cells. (A) BM-MSCs exhibited a long, spindle form and showed a whirlpool-shaped growth pattern. (B) Characterization of cell surface markers in BM-MSCs at passage 3 by flow cytometry. BM-MSCs were positive for CD105, CD90, and CD29 but negative for CD31, CD34, and CD45

Article Snippet: After 10 days of culture in DMEM (uninduced group) or inducing medium (induced group), cells were seeded into 24-well chambers, washed with PBS, fixed with 4% paraformaldehyde for 15 min, and permeabilized with 0.5% Triton X-100 for 30 min. After 1 h of blocking with 3% bovine serum albumin (BSA), cells were incubated with rabbit anti-CD31 (1:20, Abcam, Cambridge, UK) and rabbit anti- CD34 (ABclonal, Wuhan, China) antibodies in a humidified chamber at 4 °C overnight.

Techniques: Isolation, Derivative Assay, Flow Cytometry

Journal: Clinical and Applied Thrombosis/Hemostasis

Article Title: Transcriptome-wide m6A Methylation Profiling Reveals Regulatory Networks During Endothelial Differentiation of Bone Marrow Mesenchymal Stem Cells

doi: 10.1177/10760296261472670

Figure Lengend Snippet: Endothelial differentiation of BM-MSCs is confirmed by phenotypic and functional assays. (A) Flow cytometry showed an increased CD31+ cell proportion and CD34+ cell proportion after induction. (B) Western blot revealed that the expression of VE-cadherin increased after 10 days induction. (C) CD31 and CD34 immunostaining of induced cells and uninduced cells. Nuclei were stained with DAPI (blue). The expression of CD31 and CD34 were detected in induced groups. Scale bar = 200um. (D) Induced MSCs take up acetylated LDL. Scale bar = 200 um. Blue fluorescence signifies DAPI and red indicates DiI-labeled-acetylated LDL uptake in cells. (E) Tube formation assay: induced MSCs form a robust capillary-like network on Matrigel after 6 h. Scale bar = 1 mm

Article Snippet: After 10 days of culture in DMEM (uninduced group) or inducing medium (induced group), cells were seeded into 24-well chambers, washed with PBS, fixed with 4% paraformaldehyde for 15 min, and permeabilized with 0.5% Triton X-100 for 30 min. After 1 h of blocking with 3% bovine serum albumin (BSA), cells were incubated with rabbit anti-CD31 (1:20, Abcam, Cambridge, UK) and rabbit anti- CD34 (ABclonal, Wuhan, China) antibodies in a humidified chamber at 4 °C overnight.

Techniques: Functional Assay, Flow Cytometry, Western Blot, Expressing, Immunostaining, Staining, Fluorescence, Labeling, Tube Formation Assay

Journal: Frontiers in Pharmacology

Article Title: Isoalantolactone inhibits pancreatic ductal adenocarcinoma progression via direct targeting of NLRP3-mediated inflammation-angiogenesis axis

doi: 10.3389/fphar.2026.1840118

Figure Lengend Snippet: IATL demonstrates potential to inhibit PDAC angiogenesis. (A) Venn diagram illustrating the intersection between IATL-related targets and PDAC-related targets. (B) Protein–protein interaction (PPI) network of IATL targets in PDAC. (C) GO enrichment analysis of biological processes. IATL is involved in the angiogenesis process, with 5 enriched GO terms shown in the histogram. (D) KEGG pathway enrichment analysis showing that IATL participates in the classical VEGF angiogenesis pathway. The bubble plot displays 5 enriched pathways. (E,F) Laser speckle contrast imaging (LSCI) was used to evaluate blood perfusion in pancreatic cancer models. Data are presented as mean ± SD (n = 5). (G–I) Western blot analysis of CD34 and VEGFA protein levels in tumor specimens. Band intensities were quantified using ImageJ software. Data are presented as mean ± SD (n = 3). (J,K) Representative immunofluorescence images of CD34 (purple) and VEGFA (red) staining in tumor tissues from the control and IATL (5 or 10 mg/kg) treatment groups. Nuclei were counterstained with DAPI (blue). Scale bar, 50 μm. (L,M) Representative immunofluorescence images of CD34 (purple) and VEGFA (red) staining in PANC-1 cells treated with IATL (0 or 40 μM) for 24 h. Nuclei were counterstained with DAPI (blue). Scale bar, 50 μm. (N,O) Representative images of wound-healing assays in HUVECs co-cultured with conditioned medium. Images were acquired using the Cytation 7 live-cell imaging system. Wound area was quantified using ImageJ software, and the migration rate was expressed as a percentage of the control. Data are presented as mean ± SD (n = 3). (P,Q) Tube formation assays of HUVECs co-cultured with conditioned medium. Tube formation was quantified using ImageJ software. Data are presented as mean ± SD (n = 3). Statistical differences were determined by one-way ANOVA, where ns indicates no significant difference, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: Primary antibodies against VEGFA (Catalog No. A0280), IL-1β (Catalog No. A16288), IL-18 (Catalog No. A1115), ASC (CatalogNo.A1170SP)and CD34 (Catalog No. A19015) were purchased from ABclonal (China); anti-GAPDH (Catalog No.20536), GSDMD (Catalog No.20770-1-AP) and caspase (Catalog No.81482-1-RR) were sourced from proteintech (China).

Techniques: Imaging, Western Blot, Software, Immunofluorescence, Staining, Control, Cell Culture, Live Cell Imaging, Migration

Journal: Frontiers in Pharmacology

Article Title: Isoalantolactone inhibits pancreatic ductal adenocarcinoma progression via direct targeting of NLRP3-mediated inflammation-angiogenesis axis

doi: 10.3389/fphar.2026.1840118

Figure Lengend Snippet: NLRP3-mediated IATL regulates the angiogenic capacity of pancreatic cancer in vitro . (A–L) Western blot analysis of VEGFA and CD34 protein expression in PANC-1 and SW1990 cells following NLRP3 silencing or overexpression combined with IATL treatment. Band intensities were quantified using ImageJ software. Data are presented as mean ± SD (n = 3). (M–R) Tube formation assays and quantification of total vascular branch points in HUVECs cultured with pancreatic cancer cell-conditioned medium following NLRP3 intervention combined with IATL treatment. Data are presented as mean ± SD (n = 3). Statistical differences were determined by one-way ANOVA, where ns indicates no significant difference, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: Primary antibodies against VEGFA (Catalog No. A0280), IL-1β (Catalog No. A16288), IL-18 (Catalog No. A1115), ASC (CatalogNo.A1170SP)and CD34 (Catalog No. A19015) were purchased from ABclonal (China); anti-GAPDH (Catalog No.20536), GSDMD (Catalog No.20770-1-AP) and caspase (Catalog No.81482-1-RR) were sourced from proteintech (China).

Techniques: In Vitro, Western Blot, Expressing, Over Expression, Software, Cell Culture

Journal: Frontiers in Pharmacology

Article Title: Isoalantolactone inhibits pancreatic ductal adenocarcinoma progression via direct targeting of NLRP3-mediated inflammation-angiogenesis axis

doi: 10.3389/fphar.2026.1840118

Figure Lengend Snippet: IATL inhibits angiogenesis in orthotopic tumor-bearing mice by suppressing NLRP3. (A,B) Laser speckle contrast imaging (LSCI) was used to evaluate blood perfusion in pancreatic cancer models. Data are presented as mean ± SD (n = 5). (C) Representative immunofluorescence images of CD34 (purple) staining in tumor tissues from each group. Nuclei were counterstained with DAPI (blue). Scale bar, 50 μm. (D–F) Western blot analysis evaluating the effects of in vivo NLRP3 overexpression on the expression of the angiogenesis markers CD34 and VEGFA. Band intensities were quantified using ImageJ software. Data are presented as mean ± SD (n = 3). Statistical differences were determined by one-way ANOVA, where ns indicates no significant difference, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Article Snippet: Primary antibodies against VEGFA (Catalog No. A0280), IL-1β (Catalog No. A16288), IL-18 (Catalog No. A1115), ASC (CatalogNo.A1170SP)and CD34 (Catalog No. A19015) were purchased from ABclonal (China); anti-GAPDH (Catalog No.20536), GSDMD (Catalog No.20770-1-AP) and caspase (Catalog No.81482-1-RR) were sourced from proteintech (China).

Techniques: Imaging, Immunofluorescence, Staining, Western Blot, In Vivo, Over Expression, Expressing, Software