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: Cancer Cell

Article Title: Single-cell analysis defines a pancreatic fibroblast lineage that supports anti-tumor immunity

doi: 10.1016/j.ccell.2021.06.017

Figure Lengend Snippet:

Article Snippet: Anti-mouse PDGFRB (151Eu) clone APB5 , Fluidigm , 3151017B.

Techniques: Blocking Assay, Virus, Recombinant, Plasmid Preparation, Saline, Lysis, Staining, Reverse Transcription, In Vivo, Electroporation, Mass Cytometry, Conjugation Assay, Illumina Sequencing, Library Quantification, In Vitro, Quantitative RT-PCR, Software, Real-time Polymerase Chain Reaction, Cytometry, Microscopy, Imaging, Spectrophotometry

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: Cancer Research

Article Title: Discovery and Evaluation of Biomarkers for Triple-Negative Breast Cancer Subtypes Uncovers Patient Stratification and Targeted Therapeutic Strategies

doi: 10.1158/0008-5472.CAN-24-2758

Figure Lengend Snippet: TAGL as a predictive marker for dasatinib sensitivity. A, Western blot of control and TAGLN -KO cells in Hs578T (top) and BT-549 (bottom) cells. B, Representative immunofluorescence of Hs578T and BT549 control and TAGLN -KO cells, stained with αTAGL (green) and DAPI (blue). Scale bar, 50 μm. C, Cell proliferation in control (black) and TAGLN -KO (purple) cells measured by MTT assays over 72 hours. D, Relative percentage of migrated cells in control and TAGLN -KO cells, assessed by transwell assays. In C and D , values represent the mean ± SEM of at least three experimental replicates, and statistical significance was determined using one-way ANOVA. E, Drug–response curves for cell viability of control and TAGLN -KO cells treated with dasatinib at increasing concentrations. F, Western blot of known dasatinib targets in control and TAGLN -KO cells. G, Data from the DepMap portal showing the correlation between TAGLN and PDGFRB in breast cancer cell lines. H, Western blot of PDGFRβ in TAGLN -KO and cells and clones constitutively expressing PDGFRB . I, Drug–response curves for cell viability of TAGLN -KO and TAGLN -KO/ PBGFRB cells treated with dasatinib at increasing concentrations. In E and I , solid lines represent the mean of three biological replicates performed in technical replicates. The dashed line indicates their IC 50 value. **, P < 0.01; ****, P < 0.0001; ns, not significant.

Article Snippet: For PDGFRB overexpression, lentiviral particles were generated using the PDGFRB Human Tagged Lenti ORF Clone (RC206377L4, OriGene) plasmid and used to transduce target cells.

Techniques: Marker, Western Blot, Control, Immunofluorescence, Staining, Clone Assay, Expressing

Journal: Journal of Neuroinflammation

Article Title: Interferon-γ blocks signalling through PDGFRβ in human brain pericytes

doi: 10.1186/s12974-016-0722-4

Figure Lengend Snippet: Antibodies and dilutions used in this study

Article Snippet: CD140b (PDGFRβ-N-terminal) , AbD Serotec , 7460-3104 , , 1/200.

Techniques:

Journal: Journal of Cancer

Article Title: P68 RNA Helicase facilitates Breast Cancer progression by promoting Proliferation and Migration via PDGFR-β/AR axis.

doi: 10.7150/jca.61505

Figure Lengend Snippet: Figure 3. P68 RNA helicase transcriptionally regulates PDGFR-β expression in breast cancer cells. (A, B) Levels of p-PDGFRβ (IB: p-PDGFRβ) and pERK ½ (IB:pERK1/2), an established effector of PDGFR-β signaling in MDAMB231 (A) and BT549 (B) cells were analyzed by immunoblot upon PDGF-BB (20 ng/ml, 24h) stimulation. Levels of ERK ½ (IB:ERK1/2) and PDGFR-β (IB: PDGFRβ) are the loading controls. (C, D) Immunoblot analysis (C) of PDGFR-β (IB: PDGFR−β), EGFR (IB:EGFR), and FGFR (IB:FGFR) and their relative mRNA expression (D) in PDGF-BB stimulated MDAMB231 and BT549 cells upon p68 knockdown. Knockdown efficiency of p68 (IB:p68) was analyzed by immunoblot. Immunoblot of β-actin (IB:β-actin) is a loading control. (E) In vitro scratch wound healing assay of MDAMB231 cells transfected with p68 siRNA or co-transfected with p68 siRNA and PDGFRβ plasmid compared with control siRNA at 0 h and 24 h upon PDGF-BB stimulation. (F & G) Quantitative analyses of the migrating PDGF-BB stimulated MDAMB231 (F), and BT549 (G). The results represent findings of five independent experiments. Error bars represent mean ± S.E.M. **P<0.01, ****P<0.0001, ns denotes non-significant.

Article Snippet: To overexpress PDGFR−β in MDAMB231 and BT549 cells, 2.5 μg of PDGFR-β plasmid (Origene RC206377) was transfected for 48 h using LipofectamineTM 3000 Transfection Reagent kit (Thermo Fisher scientific).

Techniques: Expressing, Western Blot, Knockdown, Control, In Vitro, Wound Healing Assay, Transfection, Plasmid Preparation

Journal: Journal of Cancer

Article Title: P68 RNA Helicase facilitates Breast Cancer progression by promoting Proliferation and Migration via PDGFR-β/AR axis.

doi: 10.7150/jca.61505

Figure Lengend Snippet: Figure 5. PDGFR-β regulates AR expression and mediates the effect of DHT in proliferation of breast cancer cells. (A & B) Immunoblot analysis (A), and relative mRNA expression (B) of AR in PDGF-BB stimulated MDAMB231 and BT549 cells upon PDGFR-β knockdown. Knockdown efficiency of PDGFR-β (IB: PDGFR-β) was analyzed by immunoblot. β-actin (IB: β-actin) is the loading control. (C) Cell proliferation of MDAMB231 and BT549 cells untreated (open bar) or treated (filled bar) with DHT (1 nM for 24 h) upon PDGFR-β knock down compared with the control. The experiments were performed in triplicate. Error bars represent mean ± S.E.M. **P<0.01, ***P<0.001, ns denotes non-significant.

Article Snippet: To overexpress PDGFR−β in MDAMB231 and BT549 cells, 2.5 μg of PDGFR-β plasmid (Origene RC206377) was transfected for 48 h using LipofectamineTM 3000 Transfection Reagent kit (Thermo Fisher scientific).

Techniques: Expressing, Western Blot, Knockdown, Control

Journal: Journal of Cancer

Article Title: P68 RNA Helicase facilitates Breast Cancer progression by promoting Proliferation and Migration via PDGFR-β/AR axis.

doi: 10.7150/jca.61505

Figure Lengend Snippet: Figure 6. Nuclear PDGFR-β regulates AR expression in breast cancer cells. (A) Representative images of immunofluorescence staining of PDGFR-β (red) MDAMB231 cells treated with PDGFR-β inhibitor, AG1296 with or without PDGF-BB stimulation for 24 h compared with the control. Nuclei were stained with Hoescht (blue). (B) Levels of AR (IB: AR) in MDMB231 cells upon indicated conditions were analyzed by immunoblot. Levels of pPDGFR-β (IB: pPDGFR-β), PDGFR-β (IB:PDGFR-β) and p68 (IB:p68) are the controls, indicating the efficiency of AG1296, PDGFR-β knockdown efficiency, and total p68 respectively. Immunoblot of β-actin (IB:β-actin) is the loading control. (C) Schematic illustration of p68- PDGFRβ-AR axis in breast cancer cells upon PDGF-BB stimulation.

Article Snippet: To overexpress PDGFR−β in MDAMB231 and BT549 cells, 2.5 μg of PDGFR-β plasmid (Origene RC206377) was transfected for 48 h using LipofectamineTM 3000 Transfection Reagent kit (Thermo Fisher scientific).

Techniques: Expressing, Immunofluorescence, Staining, Control, Western Blot, Knockdown