Journal: Acta Neuropathologica Communications

Article Title: Brain-derived neurotrophic factor supports pericyte and vascular homeostasis in the aging brain

doi: 10.1186/s40478-025-02181-y

Figure Lengend Snippet: Aging reduces vasculature and pericyte expression of PDGFRβ in the brain. Brain tissue from C57BL/6J mice at 6, 12 and 24 months of age was stained for collagen type IV ( A) and quantified for total length and branch points of microvessels ( B – E ; t -test, n = 7 and 4–5 for the 6–12 months and 24 months groups, respectively). Cerebral microvessels were isolated from 12 to 24-month-old mice and homogenized for Western blot analysis of pericyte markers PDGFRβ and CD13, and protein levels of phosphorylated and total Akt and Erk1/2 ( F – L ; t -test, n = 8—9 and 7—8 per group for the 6–12 months and 24 months groups, respectively). In following experiments, transcriptomic analysis was performed using a single-cell sequencing dataset. Volcano plots show up- and down-regulated DEGs in the brains of 18-month-old mice compared with 2-month-old mice ( M ; NS, genes with non-significant changes). The bar chart displays the top five significantly enriched KEGG pathways ranked by − log 10 ( p -value) (bar length). The size of adjacent circles corresponds to the number of DEGs in each pathway. KEGG pathway analysis indicated that the down-regulated genes are associated with PI3K-AKT signaling pathway ( N )

Article Snippet: After blocking with 50 μg/ml CD16/CD32 antibody (clone 2.4G2; BioXCell, Lebanon, USA), brain cells were stained with PE-conjugated rat monoclonal antibody against mouse PDGFRβ (clone APB5; Miltenyi Biotec GmbH).

Techniques: Expressing, Staining, Isolation, Western Blot, Sequencing

Journal: Acta Neuropathologica Communications

Article Title: Brain-derived neurotrophic factor supports pericyte and vascular homeostasis in the aging brain

doi: 10.1186/s40478-025-02181-y

Figure Lengend Snippet: Deficiency of neuronal BDNF reduces vasculature and pericytes in the brain. Ten-month-old C57BL/6 mice with (ko) and without (wt) knockout of Bdnf gene in neurons for 3 months were stained for collagen type IV and quantified for the vasculature ( A ). Deficiency of neuronal BDNF significantly reduced the total length of cerebral vessels and tended to decrease the density of branch points of vessels ( B and C ; t -test, n = 6–7 per group). The brain sections were also co-stained for PDGFRβ and CD31. PDGFRβ-positive pericytes were counted and adjusted by the length of CD31-positive vessels ( D ). Deficiency of neuronal BDNF significantly reduced the number of pericytes ( E ; t -test, n = 4 per group). Additionally, microvessels were isolated from brains and detected with Western blot for pericyte markers and relevant signaling molecules ( F and I ). Deficiency of neuronal BDNF decreased protein levels of PDGFRβ but not CD13 ( G and H ; t -test, n = 6 per group). BDNF deficiency also reduced the phosphorylation of Akt, but not Erk1/2 ( J and K ; t -test, n = 4–9 per group)

Article Snippet: After blocking with 50 μg/ml CD16/CD32 antibody (clone 2.4G2; BioXCell, Lebanon, USA), brain cells were stained with PE-conjugated rat monoclonal antibody against mouse PDGFRβ (clone APB5; Miltenyi Biotec GmbH).

Techniques: Knock-Out, Staining, Isolation, Western Blot, Phospho-proteomics

Journal: Acta Neuropathologica Communications

Article Title: Brain-derived neurotrophic factor supports pericyte and vascular homeostasis in the aging brain

doi: 10.1186/s40478-025-02181-y

Figure Lengend Snippet: Deficiency of astrocyte BDNF reduces vasculature and pericytes in the brain. Brain homogenates from 10 month-old C57BL/6 mice with (ko) and without (wt) knockout of Bdnf gene in astrocytes for 3 months were detected for protein levels of BDNF ( A ). Knockout of Bdnf gene significantly reduced mature BDNF but not pro-BDNF ( B and C ; t -test, n = 9–11 per group). Brain sections were then stained for collagen type IV and quantified for the vasculature ( D ). Deficiency of astrocyte BDNF significantly reduced both the length and density of branch points of cerebral vessels (E and F; t -test, n = 8 per group). Additionally, microvessels were isolated from brains and detected with Western blot for pericyte markers and relevant signaling molecules ( G and J ). Deficiency of astrocyte BDNF decreased the protein level of PDGFRβ, but not CD13 ( H and I ; t -test, n = 11–12 per group), and reduced the phosphorylation of both Akt and Erk1/2 ( K and L ; t -test, n = 4–9 per group)

Article Snippet: After blocking with 50 μg/ml CD16/CD32 antibody (clone 2.4G2; BioXCell, Lebanon, USA), brain cells were stained with PE-conjugated rat monoclonal antibody against mouse PDGFRβ (clone APB5; Miltenyi Biotec GmbH).

Techniques: Knock-Out, Staining, Isolation, Western Blot, Phospho-proteomics

Journal: Acta Neuropathologica Communications

Article Title: Brain-derived neurotrophic factor supports pericyte and vascular homeostasis in the aging brain

doi: 10.1186/s40478-025-02181-y

Figure Lengend Snippet: BDNF acts directly on cultured pericytes. Human pericyte and endothelial cell lines were cultured. The transcription levels of BDNF receptor genes, NTRK2 and NGFR , were measured with real-time PCR. Both cells expressed NGFR gene at a significantly higher level than NTRK2 gene ( A and B ; t -test, n = 4 per group. Four experiments were independently repeated). Western blot showed expression of TrkB in the pericyte cell line ( C ). Cultured pericytes were then treated with BDNF at 0, 10, 50, and 100 ng/ml for 24 h. Western blot was used to detect the protein level of PDGFRβ, showing that BDNF treatments significantly increase PDGFRβ expression ( D and E ; One-way ANOVA followed by Bonferroni post hoc test, n = 10 per group for 0, 10 and 50 concentrations and n = 5 for 100 ng/ml concentration. Ten experiments were independently repeated), as well as to determine the phosphorylation levels of both Akt and Erk1/2 ( D , F and G ; One-way ANOVA followed by Tukey post hoc test, n = 4 or 3 per group. Four and three experiments were independently repeated for Akt and Erk1/2, respectively). Pericytes were also treated with BDNF at different concentrations in the presence of 1 µM Akt inhibitor VIII. The protein level of PDGFRβ as detected by Western blot was not altered by the treatments of BDNF ( H and I ; One-way ANOVA, p > 0.05, n = 4 per group. Four experiments were independently repeated). In additional experiments, endothelial cells were treated with BDNF at 0, 50 and 100 ng/ml for 24 h. BDNF significantly up-regulates transcription of PDGF-B , but not CD62p and CD31 genes ( J – L ; One-way ANOVA followed by Tukey post hoc test, n = 4 per group. Four experiments were independently repeated)

Article Snippet: After blocking with 50 μg/ml CD16/CD32 antibody (clone 2.4G2; BioXCell, Lebanon, USA), brain cells were stained with PE-conjugated rat monoclonal antibody against mouse PDGFRβ (clone APB5; Miltenyi Biotec GmbH).

Techniques: Cell Culture, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Concentration Assay, Phospho-proteomics

Journal: Cancers

Article Title: Tumor-Associated Fibroblast-Derived Exosomal circDennd1b Promotes Pituitary Adenoma Progression by Modulating the miR-145-5p/ONECUT2 Axis and Activating the MAPK Pathway

doi: 10.3390/cancers15133375

Figure Lengend Snippet: TAFs promoted PA progression by inhibiting miR-145-5p in PA cells. ( a ) Correlation of PA texture with PA size and aggressiveness (n = 242). ( b ) Immunohistochemical staining of TAFs indicators (α-SMA, PDGFRB, TAGLN) between soft and tough textual tumor (scale bar = 100 μm) (n = 49). ( c – e ) Correlation of TAFs density with PA size and aggressiveness. ( f ) Schematic diagram of extracting NFs and TAFs (figure created with Biorender.com). ( g ) Cell viability experiments of showing effect of TAFs supernatant on indicated PA cells (n = 6), PA1221 and PA0222 were primary PA cells. ( h ) Transwell assay of effect of TAFs supernatant on indicated PA cells (n = 6). ( i ) Analysis of tumor volume and tumor weight after indicated PA cells were mixed with NFs or TAFs (n = 6) (figure was created with Biorender.com). ( j ) Venn diagram of upregulated circRNAs and downregulated miRNAs in PA. ( k ) Heatmap of downregulated miRNAs in PA. ( l ) Correlation analysis of 5 miRNAs with TAF density (n = 15). ( m ) MiR-145-5p expression level after TAFs supernatant effect on PA cells (n = 3). ( n ) Cell viability experiments of effect of TAFs supernatant on PA cells transfected with miR-145-5p mimic (n = 6). ( o ) Colony formation assay of TAFs supernatant effect on PA cells transfected with miR-145-5p mimic (n = 3). ( p ) Analysis of tumor size and weight after miR-145-5p antagomir injection (5 nmol/3 days) (n = 6) (scale bar = 10 mm). The original magnification was ×200. Data were expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. Pearson correlation and linear regression were used to compare these parameters between two variables. One-way ANOVA was used for multi-sample comparison and Student’s t -test was used for comparison between two groups. Statistical significance was considered to be indicated by a value of p < 0.05. CM: conditioned medium.

Article Snippet: The tissues were blocked by 5% BSA for 1 h; then, the primary antibodies—α-SMA (Santa Cruz, CA, USA, cat: sc-32251), dilution ratio 1:200; PDGFRB (Proteintech, Wuhan, China), dilution ratio 1:50; TAGLN (Proteintech, Wuhan, China), dilution ratio 1:20, FGFR3 (Proteintech, Wuhan, China, cat: 66954-1-Ig), dilution ratio: 1:200—were applied overnight at 4 °C.

Techniques: Immunohistochemical staining, Staining, Transwell Assay, Expressing, Transfection, Colony Assay, Injection, Comparison

Journal: Stem Cells International

Article Title: Endothelial Cells Promote Migration of Mesenchymal Stem Cells via PDGF-BB/PDGFR β -Src-Akt in the Context of Inflammatory Microenvironment upon Bone Defect

doi: 10.1155/2022/2401693

Figure Lengend Snippet: Reagent information.

Article Snippet: JNJ-10198409 , PDGFR β , 5 μ M , 1 h , Pre-treat MSCs , MedChemExpress, Monmouth Junction, NJ, USA.

Techniques: Concentration Assay, Injection

Journal: Stem Cells International

Article Title: Endothelial Cells Promote Migration of Mesenchymal Stem Cells via PDGF-BB/PDGFR β -Src-Akt in the Context of Inflammatory Microenvironment upon Bone Defect

doi: 10.1155/2022/2401693

Figure Lengend Snippet: MSCs migrated toward ECs via PDGF-BB/PDGFR β in the inflammatory microenvironment. (a) Representative images of migrated hBMSCs that had received different pre-treatments or had been exposed to different inducing media. The migration capacity of hBMSCs was determined using a Transwell culture system. The quantification of migrated cells was shown as a bar graph. Scale bar, 50 μ m. ∗ P < 0.05. (b) Representative images of wound healing assays. The rate of scratch wound closure was shown as a bar graph. Scale bar, 200 μ m. ∗ P < 0.05. EGM: endothelial cell growth medium-2. EC-CM: conditioned media of endothelial cells. IEC-CM: conditioned media of endothelial cells in the context of inflammatory microenvironment; shPDGF-BB: short hairpin RNA targeting pdgfb ; shPDGFR β : shRNA targeting pdgfrb .

Article Snippet: JNJ-10198409 , PDGFR β , 5 μ M , 1 h , Pre-treat MSCs , MedChemExpress, Monmouth Junction, NJ, USA.

Techniques: Migration, shRNA

Journal: Stem Cells International

Article Title: Endothelial Cells Promote Migration of Mesenchymal Stem Cells via PDGF-BB/PDGFR β -Src-Akt in the Context of Inflammatory Microenvironment upon Bone Defect

doi: 10.1155/2022/2401693

Figure Lengend Snippet: Src and Akt functioned downstream of PDGFR β . (a) Representative images of migrated hBMSCs in Transwell culture systems. The quantification of migrated cells was shown as a bar graph. Data were compared with the group of IEC-CM from . Scale bar, 50 μ m. ∗ P < 0.05. (b) Representative images of wound healing assays. The rate of scratch wound closure was shown as a bar graph. Data were compared with the group of IEC-CM from . Scale bar, 200 μ m. ∗ P < 0.05. IEC-CM: conditioned media of endothelial cells in the context of inflammatory microenvironment; shPDGFR β : short hairpin RNA targeting pdgfrb ; shSrc: shRNA targeting src ; shAkt: shRNA targeting akt .

Article Snippet: JNJ-10198409 , PDGFR β , 5 μ M , 1 h , Pre-treat MSCs , MedChemExpress, Monmouth Junction, NJ, USA.

Techniques: shRNA

Journal: Stem Cells International

Article Title: Endothelial Cells Promote Migration of Mesenchymal Stem Cells via PDGF-BB/PDGFR β -Src-Akt in the Context of Inflammatory Microenvironment upon Bone Defect

doi: 10.1155/2022/2401693

Figure Lengend Snippet: Src bridged connection between PDGFR β and Akt during ECs-induced MSCs migration. (a) Gene and protein expression of Src in migrating hBMSCs. (b) Gene and protein expression of Akt in migrating hBMSCs. EC-CM: conditioned media of ECs; IEC-CM: conditioned media of ECs in the context of inflammatory microenvironment; shPDGFR β : short hairpin RNA targeting pdgfrb ; shSrc: shRNA targeting src ; shAkt: shRNA targeting akt . ∗ P < 0.05.

Article Snippet: JNJ-10198409 , PDGFR β , 5 μ M , 1 h , Pre-treat MSCs , MedChemExpress, Monmouth Junction, NJ, USA.

Techniques: Migration, Expressing, shRNA

Journal: Journal of Translational Medicine

Article Title: Papillary thyroid cancer organoids harboring BRAF V600E mutation reveal potentially beneficial effects of BRAF inhibitor-based combination therapies

doi: 10.1186/s12967-022-03848-z

Figure Lengend Snippet: List of drugs used in this study

Article Snippet: VEGFR, PDGFRβ , Lenvatinib , MedChemExpress , HY-10981 , 10.

Techniques: DNA Synthesis