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: The journal of physiological sciences : JPS

Article Title: Telocytes response to cardiac growth induced by resistance exercise training and endurance exercise training in adult male rats.

doi: 10.1186/s12576-023-00868-2

Figure Lengend Snippet: Fig. 12 CD34/PDGFR-α levels in studied groups. (*: significantly different from the control group. CTRL control group, ET endurance training group, RT: resistance training group)

Article Snippet: The rabbit polyclonal antibody to PDGFR-α (Biorbyt, UK), rabbit polyclonal antibody to PDGFR-β (Biorbyt, UK) and rabbit polyclonal antibody to CD34 (Biorbyt, UK) were added to the sample and incubated in a refrigerator at 2–8 °C for overnight in a humid environment to prevent the tissue from drying out.

Techniques: Control

Journal: The journal of physiological sciences : JPS

Article Title: Telocytes response to cardiac growth induced by resistance exercise training and endurance exercise training in adult male rats.

doi: 10.1186/s12576-023-00868-2

Figure Lengend Snippet: Fig. 13 Changes in telocytes number in response to exercise. Double-immunofluorescence labeling for CD34/PDGFR-α used to determine telocytes: these cells are increased in response to 8 weeks of endurance training (ET) and resistance training (RT)

Article Snippet: The rabbit polyclonal antibody to PDGFR-α (Biorbyt, UK), rabbit polyclonal antibody to PDGFR-β (Biorbyt, UK) and rabbit polyclonal antibody to CD34 (Biorbyt, UK) were added to the sample and incubated in a refrigerator at 2–8 °C for overnight in a humid environment to prevent the tissue from drying out.

Techniques: Immunofluorescence, Labeling

Journal: The journal of physiological sciences : JPS

Article Title: Telocytes response to cardiac growth induced by resistance exercise training and endurance exercise training in adult male rats.

doi: 10.1186/s12576-023-00868-2

Figure Lengend Snippet: Fig. 15 Changes in telocytes number in response to exercise. Double-immunofluorescence labeling for CD34/PDGFR-β used to determine telocytes: these cells are increased in response to 8 weeks of endurance training (ET) and resistance training (RT)

Article Snippet: The rabbit polyclonal antibody to PDGFR-α (Biorbyt, UK), rabbit polyclonal antibody to PDGFR-β (Biorbyt, UK) and rabbit polyclonal antibody to CD34 (Biorbyt, UK) were added to the sample and incubated in a refrigerator at 2–8 °C for overnight in a humid environment to prevent the tissue from drying out.

Techniques: Immunofluorescence, Labeling

Journal: The journal of physiological sciences : JPS

Article Title: Telocytes response to cardiac growth induced by resistance exercise training and endurance exercise training in adult male rats.

doi: 10.1186/s12576-023-00868-2

Figure Lengend Snippet: Fig. 14 CD34/PDGFR-β levels in studied groups. (*: significantly different from the control group. CTRL control group, ET endurance training group, RT resistance training group)

Article Snippet: The rabbit polyclonal antibody to PDGFR-α (Biorbyt, UK), rabbit polyclonal antibody to PDGFR-β (Biorbyt, UK) and rabbit polyclonal antibody to CD34 (Biorbyt, UK) were added to the sample and incubated in a refrigerator at 2–8 °C for overnight in a humid environment to prevent the tissue from drying out.

Techniques: Control

Journal: Biology of reproduction

Article Title: Prenatal exposure to estrogenic compounds alters the expression pattern of platelet-derived growth factor receptors alpha and beta in neonatal rat testis: identification of gonocytes as targets of estrogen exposure.

doi: 10.1095/biolreprod.102.009605

Figure Lengend Snippet: FIG. 4. Quantification of in situ hybridization signals for PDGFRa and b mRNAs in 3-day-old testis sections. Densitometric analysis was per- formed as described in the Materials and Methods section. The histograms represent the specific intensities of signals, expressed in arbitrary units, for PDGFRa (A) and PDGFRb (B) that were present in the interstitium, periphery, and center of the seminiferous cords of the same samples as Figure 3. *P , 0.05; ***P , 0.001.

Article Snippet: Rabbit polyclonal antibodies against the Cterminal domains of PDGFRa and PDGFRb were purchased from Santa Cruz Biotechnology (Santa Cruz, CA), while rabbit polyclonal antibodies against the kinase domain of PDGFRb was purchased from BD Biosciences (BD Pharmingen, San Diego, CA).

Techniques: In Situ Hybridization

Journal: Diagnostic Pathology

Article Title: Diagnostic potential of NRG1 in benign nerve sheath tumors and its influence on the PI3K-Akt signaling and tumor immunity

doi: 10.1186/s13000-024-01438-9

Figure Lengend Snippet: Detection of NRG1, immune cells influenced by NRG1, and PI3K/AKT in clinical tissue samples. A IHC detection of S100 expression in clinical tissue samples. con: normal vestibular tissue. S100 is mainly expressed in the BNST cytoplasm. S100 is a biological marker of BNST—scale bar: 20 μm. The statistical graph is on the right. B RT-qPCR detection of Nrg1 mRNA expression levels in clinical tissue samples. C Western blot detection of NRG1 and key proteins in the PI3K/AKT pathway in clinical tissue samples. The statistical graph is on the right. D IHC detection of NRG1 expression in clinical tissue samples. Mainly distributed in the cytoplasm and membrane of myelin-forming cells and macrophages. Scale bar: 20 μm. The statistical graph is on the right. E Changes in M2 macrophages (M2 mac) and neutrophils (neu) in clinical samples detected by IHC. CD206-scale bar: 20 μm. CD66b: scale bar: 20 μm. Both are expressed on the cell membrane of macrophages and neutrophils. Statistical graphs on the right. F Changes in M2 macrophages and neutrophils in clinical samples detected by flow cytometry. M2 mac: CD68 + CD206 + , neu: CD14 + CD66b + . Statistical graph in the first quadrant. con group: n = 3; BNST group: n = 5. Indicates a difference compared to con, where P < 0.05 indicates a difference compared to con, P < 0.01, and *** indicates a difference compared to con, P < 0.0001

Article Snippet: Following cell infection, a treatment involving 25 μg/ml of the PI3K activator 740 Y-P (HY-P0175, MedChemExpress) was administered for a duration of 48 h [ ].

Techniques: Expressing, Marker, Quantitative RT-PCR, Western Blot, Membrane, Flow Cytometry

Journal: Diagnostic Pathology

Article Title: Diagnostic potential of NRG1 in benign nerve sheath tumors and its influence on the PI3K-Akt signaling and tumor immunity

doi: 10.1186/s13000-024-01438-9

Figure Lengend Snippet: Validation of NRG1-mediated cell behavior through the PI3K/AKT signaling pathway using cell line models. A RT-qPCR detection of NRG1 knockdown efficiency. B Western blot detection of NRG1 knockdown efficiency and expression levels of critical proteins in the PI3K/AKT signaling pathway. Statistical graphs on the right. C Western blot detection of critical proteins in the PI3K/AKT signaling pathway in cells treated with a PI3K inhibitor. Statistical graphs on the right. D Cell cycle of cells after NRG1 knockdown and PI3K inhibitor treatment detected by flow cytometry. Statistical graphs on the right. E After NRG1 knockdown and PI3K inhibitor treatment, apoptosis proportions of cells were detected by flow cytometry. Statistical graph in the first quadrant. F Cell migration ability after NRG1 knockdown and PI3K inhibitor treatment detected by Transwell assay. G Expression levels of critical proteins in the PI3K/AKT signaling pathway in cells treated with a PI3K agonist after NRG1 knockdown detected by Western blot. Statistical graphs on the right. H Cell cycle changes of cells after NRG1 knockdown and PI3K agonist treatment detected by flow cytometry. Statistical graphs on the right. I After NRG1 knockdown and PI3K agonist treatment, apoptosis of cells was detected by flow cytometry. Statistical graph in the first quadrant. J Cell migration ability after NRG1 knockdown and PI3K agonist treatment detected by Transwell assay. Indicates a difference compared to sh-NC or con or sh-Nrg1 + DMSO, where P < 0.05, indicates a difference compared to sh-NC or DMSO or sh-Nrg1 + DMSO, P < 0.01, and indicates a difference compared to sh-NC or DMSO or sh-Nrg1 + DMSO, P < 0.001, ** indicates a difference compared to sh-NC or DMSO or sh-Nrg1 + DMSO, P < 0.0001

Article Snippet: Following cell infection, a treatment involving 25 μg/ml of the PI3K activator 740 Y-P (HY-P0175, MedChemExpress) was administered for a duration of 48 h [ ].

Techniques: Biomarker Discovery, Quantitative RT-PCR, Knockdown, Western Blot, Expressing, Flow Cytometry, Migration, Transwell Assay

Journal: Diagnostic Pathology

Article Title: Diagnostic potential of NRG1 in benign nerve sheath tumors and its influence on the PI3K-Akt signaling and tumor immunity

doi: 10.1186/s13000-024-01438-9

Figure Lengend Snippet: Validation of NRG1-mediated BNST progression through the PI3K/AKT signaling pathway using PDX models. A In vivo fluorescence imaging representation of tumor growth after sciatic nerve heterotopic transplantation. B Changes in critical proteins in the PI3K/AKT signaling pathway detected by Western blot in xenografts. C Statistical graph of protein expression in Figure B. C Tumor growth curve of PDX mice. D Kaplan-Meier survival curve of PDX mice. F Representative images of Ki67 IHC staining in xenografts. Scale bar: 100 μm. Ki67 is mainly expressed in cell nuclei. G Apoptosis of cells in xenografts detected by flow cytometry. Statistical graph in the first quadrant. H Flow cytometry detected the proportion of M2 macrophages (M2 mac) and neutrophils (neu) in xenografts—statistical graph in the first quadrant. M2 mac: CD68 + CD206 + , neu: CD14 + CD66b + ; n = 3 per group. Indicates a difference compared to PDX-NC or PDX-con, where P < 0.05 indicates a difference compared to PDX-NC or PDX-con, P < 0.01 indicates a difference compared to PDX-NC or PDX-con, P < 0.001, ** indicates a difference compared to PDX-NC or PDX-con, P < 0.0001

Article Snippet: Following cell infection, a treatment involving 25 μg/ml of the PI3K activator 740 Y-P (HY-P0175, MedChemExpress) was administered for a duration of 48 h [ ].

Techniques: Biomarker Discovery, In Vivo, Fluorescence, Imaging, Transplantation Assay, Western Blot, Expressing, Immunohistochemistry, Flow Cytometry

Journal: PLoS ONE

Article Title: IFNγ-stimulated dendritic cell extracellular vesicles can be nasally administered to the brain and enter oligodendrocytes

doi: 10.1371/journal.pone.0255778

Figure Lengend Snippet: IFNγ-DC-EVs were transfected with reporter mCherry mRNA. (A) Nasal delivery of transfected EVs (NA mCherry-EV) confirmed EV delivery of functional mCherry mRNA to brain cells. Representative images show mCherry fluorescence (red), anti-mCherry immunostaining (green) and merged image (yellow) to confirm mCherry expression in cells. Animals that were nasally administered untransfected EVs (Sham) showed no mCherry expression/staining. Scale bar = 25 μm. (B) Representative merged images (B1-8) show widespread cellular distribution in the CNS two days after nasal administration of mCherry mRNA transfected IFNγ-DC-EVs. Images were taken from brain sections represented in the schematic. A subset of mCherry-positive cells colocalize with CNPase, indicating that they are oligodendrocytes. (C) Representative high power images of mCherry expression (red) in cells that co-stained (yellow) for CNPase (green). Scale bar = 25 μm. (D) Similarly, stained representative merged images (D1-8) show oligodendrocyte co-staining throughout the CNS, with numbers corresponding to brain areas in schematic (B). Scale bar = 20 μm. ( n = 5-7/group; A-D). To confirm that IFNγ-DC-EVs delivered cargo to oligodendrocytes, protein expression levels of known miR-219 mRNA targets expressed in oligodendrocyte lineage cells were quantified via immunoblot. Three days after nasal delivery of IFNγ-DC-EVs or Unstim-DC-EVs [Sham(S)], levels of (E) PDGFRα, and (F) ELOVL7 were significantly (* p = 0.040 and 0.038 respectively) reduced in the brains of animals that received IFNγ-DC-EVs. ( n = 3-6/group).

Article Snippet: Primary antibodies used were PDGFRα (#NBP2-67025; Novus Biologicals) at 1:5000, and ELOVL7 (#NBP1-93926; Novus Biologicals) at 1:750.

Techniques: Transfection, Functional Assay, Fluorescence, Immunostaining, Expressing, Staining, Western Blot