Journal: Journal of Biomedical Science

Article Title: Many but not all pathogen-associated molecular patterns aggravate neurogenic heterotopic ossification after spinal cord injury

doi: 10.1186/s12929-026-01237-y

Figure Lengend Snippet: Expression of PRRs by mouse and human skeletal muscle cells. Expression of PRR mRNA transcripts in mouse satellite cells (SC), FAPs (FP), endothelial cells (EC) and monocyte/macrophages (MΦ) sorted from skeletal muscles. Whole mouse bone marrow (BM) cells and splenocytes (SP) were used as positive controls for these transcripts. Human PDGFRα + FAPs were sorted from muscles surrounding NHO biopsies, and CD14 + monocytes were isolated from peripheral blood from healthy donors. A TLR1, B TLR2, C TLR3, D TLR4, E TLR5, F TLR6, G TLR7, H TLR8, I TLR9, J STING1, K RIGI, L MDA5 (IFIH1), M PRK, N NOD1, O NOD2, P Dectin-1 (CLEC7A), Q Dectin-2 (Clec4n/CLEC6A), R Mincle (CLEC4E). For each gene, left hand histogram represents expression of the mouse gene in mouse cells and the right-hand histogram represents expression of the human gene in human cells. For mouse cells, relative mRNA expression was quantified relative to house-keeping gene Hprt . For human cells, values were normalized using three references genes HPRT , RPLP0 and PPIA for FAPs and ACTB , GAPDH and PPIA for CD14 + cells. Each dot represents a mouse or a human donor, bars represent mean ± SD

Article Snippet: Human MPCs were trypsinized and incubated for 30 min with biotinylated anti-human PDGFRα (Cat# BAF322, R&D Systems) goat polyclonal antibody and CD56-PE (clone B159, BD Pharmigen) monoclonal antibody in PBS 2% FCS, 2 mM EDTA or with control isotypes IgG1 PE (Cat# A07796, Beckman Coulter) and biotinylated goat IgG (Cat# BAF108, R&D Systems).

Techniques: Expressing, Muscles, Isolation

Journal: Journal of Biomedical Science

Article Title: Many but not all pathogen-associated molecular patterns aggravate neurogenic heterotopic ossification after spinal cord injury

doi: 10.1186/s12929-026-01237-y

Figure Lengend Snippet: OSM and IL-1 neutralization in monocyte-conditioned media strongly inhibit hFAPs mineralization. A OSM, IL-1α and IL-1β concentrations quantified in conditioned media from human CD14 + macrophages stimulated with 200 ng/ml Pam2CSK4 (CM Pam2CSK4 ), 200 ng/ml Pam3CSK4 (CM Pam3CSK4 ) or non-stimulated (CM ∅ ). B OSM, IL-1α and IL-1β concentrations in CM Pam2CSK4 , CM Pam3CSK4 and CM ∅ were correlated with hFAP calcium mineralization measured by Alizarin Red staining after 2 weeks of culture in osteogenic conditions in the presence of the conditioned media. Each dot represents a different conditioned medium sample. C–F Mouse anti-human OSM antibody, isotype control antibody and IL-1RA were used to neutralize OSM and IL-1 in human CD14 + monocyte conditioned media as indicated below each chart. Calcium mineralization of hFAPs cultured for 2 weeks in osteogenic conditions with C CM Pam2CSK4 or D CM Pam3CSK4 was measured using Alizarin Red staining and quantified by spectrophotometry. E, F RUNX2 protein quantification by Western blot using Stain-Free normalization for hFAPs cultured for 2 weeks in osteogenic conditions with E CM Pam2CSK4 or F CM Pam3CSK4 . In C-F, each dot represents a hFAP individual donor. Bars represent mean ± SD, One-way ANOVA Dunnett’s multiple comparison test versus CM ∅ in ( A ) and ( B ), versus CM Pam2CSK4 in ( C ) and ( E ), or versus CM Pam3CSK4 in ( D ) and ( F ), * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Human MPCs were trypsinized and incubated for 30 min with biotinylated anti-human PDGFRα (Cat# BAF322, R&D Systems) goat polyclonal antibody and CD56-PE (clone B159, BD Pharmigen) monoclonal antibody in PBS 2% FCS, 2 mM EDTA or with control isotypes IgG1 PE (Cat# A07796, Beckman Coulter) and biotinylated goat IgG (Cat# BAF108, R&D Systems).

Techniques: Neutralization, Staining, Control, Cell Culture, Spectrophotometry, Western Blot, Comparison

Journal: Reproduction (Cambridge, England)

Article Title: Platelet-derived growth factor modulates the primordial to primary follicle transition.

doi: 10.1530/rep.1.00978

Figure Lengend Snippet: Figure 1 Effect of PDGF-treatment on primordial to primary follicle transition in cultured ovaries. Ovaries from 4-day-old rats were placed into culture for 14 days. Cultured ovaries were treated with PDGF, anti- PDGF neutralizing antibody, KITL or were left untreated as controls. After culture all ovaries were fixed, stained and subjected to morphological analysis. The follicles per ovary cross-section were categorized as being either primordial or developing (which includes all follicles having undergone the primordial to primary transition). Data are presented as the mean percentageGS.E.M. with data pooled from four separate experiments (nZ6–12 ovaries per treatment group). One-way ANOVA showed a significant (P!0.0001) effect of treatment. Asterisks indicate that the percentage of developing follicles for a treatment is significantly (P!0.05) different than that of the control by post-hoc Dunnet’s test.

Article Snippet: Treatments during organ culture included recombinant rat PDGF-AB heterodimer (R&D Systems, Inc.; Minneapolis, MN, USA) at 50 ng/ml, rat KITL (Amgen; Thousand Oaks, CA, USA) at 50 ng/ml, anti-PDGF neutralizing antibody (R&D Systems) at 20 mg/ml, www.reproduction-online.org human NRG1-b1 extracellular domain (R&D Systems) at 50 ng/ml, human NRG1-b1 EGF domain (R&D Systems) at 50 ng/ml, or rat VEGF (R&D Systems) at 50 ng/ml.

Techniques: Cell Culture, Staining, Control

Journal: Reproduction (Cambridge, England)

Article Title: Platelet-derived growth factor modulates the primordial to primary follicle transition.

doi: 10.1530/rep.1.00978

Figure Lengend Snippet: Figure 3 ImmunohistochemicallocalizationofPDGFproteininculturedovaries.ThepresenceofPDGFproteinisindicatedbyadarkbrownstaininovary sections. A) Four-day-old ovaries cultured for 14 days showing PDGF staining at high intensity in oocytes. B) Control cultured ovary section stained using non-immune IgG as a primary antibody. C) Freshly isolated 4-day-old ovaries showing PDGF staining in the oocytes of primordial follicles. D) Control freshly isolated ovary section stained using non-immune IgG as a primary antibody. Images are representative of two different experiments using ovaries from different rats. Microscope magnificationZ400!. Total microscope magnification at the time of image capture was 400!.

Article Snippet: Treatments during organ culture included recombinant rat PDGF-AB heterodimer (R&D Systems, Inc.; Minneapolis, MN, USA) at 50 ng/ml, rat KITL (Amgen; Thousand Oaks, CA, USA) at 50 ng/ml, anti-PDGF neutralizing antibody (R&D Systems) at 20 mg/ml, www.reproduction-online.org human NRG1-b1 extracellular domain (R&D Systems) at 50 ng/ml, human NRG1-b1 EGF domain (R&D Systems) at 50 ng/ml, or rat VEGF (R&D Systems) at 50 ng/ml.

Techniques: Cell Culture, Staining, Control, Isolation, Microscopy

Journal: Reproduction (Cambridge, England)

Article Title: Platelet-derived growth factor modulates the primordial to primary follicle transition.

doi: 10.1530/rep.1.00978

Figure Lengend Snippet: Figure 5 Kit ligand (KITL) mRNA expression in cultured ovaries in response to PDGF-treatment. Ovaries from 4-day-old rats were placed into culture and treated with PDGF or were not treated for 2 days. Real- time PCR was performed on RNA isolated from cultured whole ovaries to determine levels of KITL mRNA expression. Data are expressed as KITL mRNA/S2 mRNA normalized to untreated control values. Data are from three separate experiments with the meanGS.E.M. presented. Asterisk indicates a significant difference (P!0.05) from control as determined by a one-sample t-test.

Article Snippet: Treatments during organ culture included recombinant rat PDGF-AB heterodimer (R&D Systems, Inc.; Minneapolis, MN, USA) at 50 ng/ml, rat KITL (Amgen; Thousand Oaks, CA, USA) at 50 ng/ml, anti-PDGF neutralizing antibody (R&D Systems) at 20 mg/ml, www.reproduction-online.org human NRG1-b1 extracellular domain (R&D Systems) at 50 ng/ml, human NRG1-b1 EGF domain (R&D Systems) at 50 ng/ml, or rat VEGF (R&D Systems) at 50 ng/ml.

Techniques: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Isolation, Control

Journal: Reproduction (Cambridge, England)

Article Title: Platelet-derived growth factor modulates the primordial to primary follicle transition.

doi: 10.1530/rep.1.00978

Figure Lengend Snippet: Figure 6 A model of the regulation of the primordial to primary follicle transition by paracrine growth factors, including the actions of platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), leukemia inhibitory factor (LIF), bone morphogenetic protein-4 (BMP4), keratinocyte growth factor (KGF), Anti-Mu¨llerian Hormone/Mu¨llerian inhibitory substance (AMH/MIS), and insulin.

Article Snippet: Treatments during organ culture included recombinant rat PDGF-AB heterodimer (R&D Systems, Inc.; Minneapolis, MN, USA) at 50 ng/ml, rat KITL (Amgen; Thousand Oaks, CA, USA) at 50 ng/ml, anti-PDGF neutralizing antibody (R&D Systems) at 20 mg/ml, www.reproduction-online.org human NRG1-b1 extracellular domain (R&D Systems) at 50 ng/ml, human NRG1-b1 EGF domain (R&D Systems) at 50 ng/ml, or rat VEGF (R&D Systems) at 50 ng/ml.

Techniques: Derivative Assay

Journal: The Journal of Neuroscience

Article Title: Developing Schwann Cells Acquire the Ability to Survive without Axons by Establishing an Autocrine Circuit Involving Insulin-Like Growth Factor, Neurotrophin-3, and Platelet-Derived Growth Factor-BB

doi: 10.1523/jneurosci.19-10-03847.1999

Figure Lengend Snippet: Figure 3. Dose–response curves of NT-3, IGF-2, and PDGF-BB alone and on the background of the other two factors. Note that each of these factors unambiguously promotes Schwann cell survival in a dose- dependent manner provided the other two factors are present. Note that when applied singly at the concentration used in the minimal mixture (IGF-2, 1.6 ng/ml; NT-3 and PDGF-BB, 0.8 ng/ml), the effect of each factor is small, although the mixture supports full survival (see Results and Fig. 2B, first column from left) showing a synergistic action in promoting survival. The IGF-2 curve was constructed in the presence of PDGF-BB and NT-3 both at 0.2 ng/ml. The NT-3 curve was constructed in the presence of PDGF-BB at 0.2 ng/ml and IGF-2 at 0.4 ng/ml. The PDGF-BB curve was constructed in the presence of NT-3 at 0.2 ng/ml and IGF-2 at 0.4 ng/ml. PORN substrate, 125 cells per coverslip, 2 d assay.

Article Snippet: Neutralizing polyclonal PDGF-BB antibodies were from New Brunswick Scientific (Huntingdon, UK).

Techniques: Concentration Assay, Construct

Journal: The Journal of Neuroscience

Article Title: Developing Schwann Cells Acquire the Ability to Survive without Axons by Establishing an Autocrine Circuit Involving Insulin-Like Growth Factor, Neurotrophin-3, and Platelet-Derived Growth Factor-BB

doi: 10.1523/jneurosci.19-10-03847.1999

Figure Lengend Snippet: Figure 4. b-Neuregulins support Schwann cell survival but are unlikely to be a major component of the survival signal in conditioned media. A, A soluble form of the b-neuregulin-1 receptor ErbB4 inhibits b-neuregulin-1-mediated survival but has no effect on survival mediated by the minimal mixture of IGF-2, NT-3, and PDGF-BB or conditioned medium (dilution, 1:10). Note that for direct comparison, cell number at the end of the assay (2 d) in b-neuregulin-1 alone is arbitrarily set at 100%. Cell number in the presence of b-neuregulin-1 plus added ErbB4 is normalized to this value to obtain relative survival. NRG-b1, b-Neuregulin-1, 8 ng/ml. PORN substrate, 125 cells per coverslip, 2 d assay. B, b-Neuregulin-mediated survival is unaffected by neutralizing antibodies to IGF, NT-3, or PDGF-BB. DM, Simple defined medium; NRG-b1, b-neuregulin-1, 8 ng/ml. For anti-NT-39 and anti-NT-30 see legend to Figure 2A; anti-IGF, IGF antibody SM1.2. The antibodies were applied at the same concentrations as those used to inhibit activity in conditioned media. PORN substrate, 125 cells per coverslip, 2 d assay.

Article Snippet: Neutralizing polyclonal PDGF-BB antibodies were from New Brunswick Scientific (Huntingdon, UK).

Techniques: Comparison, Activity Assay

Journal: The Journal of Neuroscience

Article Title: Developing Schwann Cells Acquire the Ability to Survive without Axons by Establishing an Autocrine Circuit Involving Insulin-Like Growth Factor, Neurotrophin-3, and Platelet-Derived Growth Factor-BB

doi: 10.1523/jneurosci.19-10-03847.1999

Figure Lengend Snippet: Figure 6. Semiquantitative RT-PCR analysis of the putative autocrine factors and their respective receptor mRNAs during development, in culture, and after nerve transection. A, RT-PCR on neonatal sciatic nerve (Ne) and Schwann cells cultured from neonatal nerves (Sch). Examination of IGF-1 (43 cycles), IGF-2 (28 cycles), PDGF-B (33 cycles), and NT-3 (48 cycles) and their corresponding receptors IGF-RI (45 cycles), IGF- RII (33 cycles), PDGF-Rb (33 cycles), and TrkC (36 cycles) reveals the presence of all mRNAs in newborn nerve as well as Schwann cells in culture. B, These mRNAs are expressed in E18 nerves (immature Schwann cells) but also earlier in embryonic development at E14 (pre- cursor stage) and E16 (a transition point between precursors and imma- ture Schwann cells). The cycle numbers for each primer pair were 33 cycles for IGF-1, 27 cycles for IGF-2, 46 cycles for NT-3, and 33 cycles for the TrkC receptor. For all other primer pairs the cycle numbers were identical to those listed in A. C, After nerve transection, mRNAs for all factors and their receptors are expressed at the levels that are broadly comparable to the contralateral control side. CON, Normal nerves of 2- and 4-d-old rats; CUT, the distal stump of 2- and 4-d-old rats 2 and 4 d after transection performed at birth. Cycle numbers were 30 for IGF-1, 27 for IGF-2, 40 for NT-3, and 35 for TrkC. For PDGF-B and PDGF-Rb 34 cycles were done. Again, 45 cycles were run for IGF-RI, and 33 were run for IGF-RII.

Article Snippet: Neutralizing polyclonal PDGF-BB antibodies were from New Brunswick Scientific (Huntingdon, UK).

Techniques: Reverse Transcription Polymerase Chain Reaction, Cell Culture, Control

Journal: The Journal of Neuroscience

Article Title: Developing Schwann Cells Acquire the Ability to Survive without Axons by Establishing an Autocrine Circuit Involving Insulin-Like Growth Factor, Neurotrophin-3, and Platelet-Derived Growth Factor-BB

doi: 10.1523/jneurosci.19-10-03847.1999

Figure Lengend Snippet: Figure 5. Survival in conditioned medium or in IGF-2, NT-3, and PDGF-BB depends on MAP kinase activation, but survival in b-neuregulin-1 is MAP kinase-independent. The graph shows the effect of blocking the MAP kinase pathway (using 20, 35, or 50 mM PD98059) on survival under three different conditions: in the minimal mixture of IGF-2, NT-3, and PDGF-BB, in conditioned medium at a dilution of 1:10 (Cond.medium), and in b-neuregulin-1 at 8 ng/ml (NRG-b1). Note that PD98059 has no effect on cell numbers in b-neuregulin. The number is higher at the end of the assay than the number of cells initially plated because of the mitogenic effect of b-neuregulin-1. The experiment was done in simple defined medium using PORN substrate, 125 cells per coverslip, and a 2 d assay.

Article Snippet: Neutralizing polyclonal PDGF-BB antibodies were from New Brunswick Scientific (Huntingdon, UK).

Techniques: Activation Assay, Blocking Assay

Journal: The Journal of Neuroscience

Article Title: Developing Schwann Cells Acquire the Ability to Survive without Axons by Establishing an Autocrine Circuit Involving Insulin-Like Growth Factor, Neurotrophin-3, and Platelet-Derived Growth Factor-BB

doi: 10.1523/jneurosci.19-10-03847.1999

Figure Lengend Snippet: Figure 9. The emergence of an autocrine survival circuit coincides with the generation of Schwann cells from precursors. A, Essentially all E14 precursors die irrespective of plating density, whereas the survival of immature Schwann cells from E18 nerves is density-dependent. Identical results were obtained on laminin or PORN substrates in experiments with precursors. E18 Schwann cells die more slowly on laminin than on PORN. Therefore, at the 2 d time points shown here, more cells are still alive at each density point on laminin compared with PORN substrates. One day assay for E14 precursors, 2 d assay for E18 Schwann cells. B, E14 Schwann cell precursors cannot be rescued by Schwann cell conditioned medium or by IGF-2, NT-3, and PDGF-BB. b-Neuregulin, an established precursor survival factor, is included as a positive control. DM, Simple defined medium; CM, Schwann cell conditioned medium at a dilution of 1:10. IGF-2, NT-3, and PDGF-BB were at the minimal concentrations, and b-neuregulin-1 was at 2 ng/ml. PORN substrate, 1 d assay. Identical results were obtained when the experiments shown in B were performed in supplemented defined medium and when the experiments were per- formed on laminin substrate (data not shown).

Article Snippet: Neutralizing polyclonal PDGF-BB antibodies were from New Brunswick Scientific (Huntingdon, UK).

Techniques: Positive Control

Journal: The Journal of Neuroscience

Article Title: Developing Schwann Cells Acquire the Ability to Survive without Axons by Establishing an Autocrine Circuit Involving Insulin-Like Growth Factor, Neurotrophin-3, and Platelet-Derived Growth Factor-BB

doi: 10.1523/jneurosci.19-10-03847.1999

Figure Lengend Snippet: Figure 8. In the presence of autocrine signals, laminin promotes long- term survival. A, In sparse cultures, Schwann cells die on laminin sub- strate. Laminin substrate, 125 cells per coverslip. B, Laminin promotes longer-term survival in the minimal mixture of IGF-2, NT-3, and PDGF-BB and supports full survival for at least 6 d in the presence of conditioned medium (CM; dilution, 1:10). Laminin substrate, 125 cells per coverslip.

Article Snippet: Neutralizing polyclonal PDGF-BB antibodies were from New Brunswick Scientific (Huntingdon, UK).

Techniques:

Journal: The Journal of Neuroscience

Article Title: Developing Schwann Cells Acquire the Ability to Survive without Axons by Establishing an Autocrine Circuit Involving Insulin-Like Growth Factor, Neurotrophin-3, and Platelet-Derived Growth Factor-BB

doi: 10.1523/jneurosci.19-10-03847.1999

Figure Lengend Snippet: Figure 10. From paracrine signals to autocrine loops: the proposed changes in survival regulation during Schwann cell development. The survival of E14 Schwann cell precursors is regulated in a paracrine manner by axon-derived b-neuregulin-1. During development, Schwann cells establish autocrine circuits, which, in neonatal nerves, act in parallel with the axonal signal. During Schwann cell maturation the autocrine signal becomes sufficient to prevent Schwann cell death in the absence of axons. The factors IGF, NT-3, and PDGF-BB have been shown to be major components of this autocrine signal in Schwann cells from 7-d-old nerves. In vivo experiments indicate that at this date loss of axonal contact after nerve transection no longer triggers significant Schwann cell death (Z. Dong, R. Mirsky, and K. R. Jessen, unpublished observations). A, Axons; P, Schwann cell precursors; S, Schwann cells.

Article Snippet: Neutralizing polyclonal PDGF-BB antibodies were from New Brunswick Scientific (Huntingdon, UK).

Techniques: Derivative Assay, In Vivo

Journal: Cell Death & Disease

Article Title: RASSF1A inhibits PDGFB-driven malignant phenotypes of nasopharyngeal carcinoma cells in a YAP1-dependent manner

doi: 10.1038/s41419-020-03054-z

Figure Lengend Snippet: a A heat map generated using the significantly changed genes categorized in the “cytokine-cytokine receptor interaction pathway” is shown. b , c mRNA expression ( b ) was evaluated by qRT-PCR and protein concentration by ELISA ( c ) in CM of RASSF1A-overexpressing CNE-2 cells, RASSF1A-depleted CNE-1 cells and their corresponding control cells, The data are presented as the mean ± S.D. values, ** p < 0.01, Student’s t test. d – g PDGFB was transiently knocked down with a pool of siRNA or treated with a neutralizing antibody for PDGF-BB (10 µg/mL) in RASSF1A-depleted CNE-1 cells. PDGF-BB secretion in the CM was measured by ELISA ( d ), ** p < 0.01, Student’s t test. e Number of spheroids formed was determined via microscopy, and representative images ( e left panel) are shown. The formed spheroids were compared ( e right panel), the data are presented as the mean ± S.D. values, * p < 0.05, ** p < 0.01, Student’s t test; ns: non-sinificant. Scale bar: 200 µm. Representative images of the migration assay ( f ) and invasion assay ( g ) are shown, the data are presented as the mean ± S.D. values, ** p < 0.01, Student’s t test. Scale bar: 100 µm. h – j Recombinant PDGF-BB or IgG was added to RASSF1A-overexpressing CNE-2 cells. Representative images of sphere formation ( h ) (Scale bar: 200 µm.), migration ( i ) and invasion ( j ) assays (Scale bar: 100 µm) of RASSF1A-overexpressing CNE-2 cells treated with PDGF-BB (the culture medium was supplemented with 20 ng/ml or an equal volume of control IgG) are shown, The data are presented as the mean ± S.D. values, * p < 0.05, ** p < 0.01, Student’s t test.

Article Snippet: Cells were plated in 6-well plates (Corning, USA) and treated with humane recombinant PDGF-BB (220-BB-010, R&D Systems, USA) or Immunoglobulin G (IgG) control (AB-108-C, R&D Systems, USA) or neutralizing antibody against PDGF-BB (AB-220-NA, R&D Systems, USA) or latrunculin b (LTB, ab144291, Abcam, UK) 12 h after plating.

Techniques: Generated, Expressing, Quantitative RT-PCR, Protein Concentration, Enzyme-linked Immunosorbent Assay, Control, Microscopy, Migration, Invasion Assay, Recombinant

Journal: Cell Death & Disease

Article Title: RASSF1A inhibits PDGFB-driven malignant phenotypes of nasopharyngeal carcinoma cells in a YAP1-dependent manner

doi: 10.1038/s41419-020-03054-z

Figure Lengend Snippet: a – c YAP1 was transiently knocked down in RASSF1A-depleted CNE-1 cells. a In the indicated cells, YAP1 protein expression was assessed by using western blotting; b PDGFB, CYP61 and CTGF mRNA expression was assessed by qRT-PCR; c Concentration of PDGF-BB secreted in CM was measured by ELISA. The data are presented as the mean ± S.D. values, * p < 0.05, ** p < 0.01, Student’s t test. d – f Recombinant PDGF-BB or IgG was added to YAP1-silenced NPC cells. d The formed spheroids were counted via microscopy, and ( e ) representative images are shown. f The impact of PDGF-BB treatment on the migration and invasion of RASSF1A-depleted cells was determined by Transwell assays. The data are presented as the mean ± S.D. values, * p < 0.05, ** p < 0.01, Student’s t test. Scale bar: 200 µm.

Article Snippet: Cells were plated in 6-well plates (Corning, USA) and treated with humane recombinant PDGF-BB (220-BB-010, R&D Systems, USA) or Immunoglobulin G (IgG) control (AB-108-C, R&D Systems, USA) or neutralizing antibody against PDGF-BB (AB-220-NA, R&D Systems, USA) or latrunculin b (LTB, ab144291, Abcam, UK) 12 h after plating.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Concentration Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Microscopy, Migration

Journal: Circulation

Article Title: Platelet-derived growth factor-AB limits the extent of myocardial infarction in a rat model: feasibility of restoring impaired angiogenic capacity in the aging heart.

doi: 10.1161/hc0502.103672

Figure Lengend Snippet: Figure 3. A, Representative vWF (DAB) staining of vessels in 4- and 24-month-old rat hearts pretreated with vehicle or PDGF-AB 24 h before euthanization (magnification 200). B, Vascular density as determined by vWF staining for the sets of treated rats (number of vessels per high-power field [HPF; mag- nification 400]) (n3 for each set). *P0.05 vehicle vs control; vascular density differences between 4- and 24-month-old treat- ment sets are not statistically significant.

Article Snippet: In addition, at the time of cardiac or pulmonary allograft transplantation, sets of young adult mice also were treated with single subcutaneous pinnal injections of antibodies to neutralize PDGF-AB (10 g in 20 L PBS, AB-20-NA, R&D Systems, n 8 cardiac, 8 pulmonary allografts), PDGFR- , PDGFR- (10 g in 20 L PBS, AB-307-NA and AB385, R&D Systems, respectively; n 7 cardiac allografts), or nonimmune rabbit immunoglobulin G (10 g in 20 L PBS, AB-105-C, R&D Systems; n 8 cardiac, 8 pulmonary allografts).

Techniques: Staining, Control

Journal: Circulation

Article Title: Platelet-derived growth factor-AB limits the extent of myocardial infarction in a rat model: feasibility of restoring impaired angiogenic capacity in the aging heart.

doi: 10.1161/hc0502.103672

Figure Lengend Snippet: Figure 4. Representative Masson’s trichrome staining in 4-month-old rat hearts pretreated with PBS or PDGF-AB 24 h before LAD ligation. Graph shows myocardial infarct size scored 14 days after coronary artery ligation (control, n13; PDGF-AB, n12). *P0.02, PDGF vs control.

Article Snippet: In addition, at the time of cardiac or pulmonary allograft transplantation, sets of young adult mice also were treated with single subcutaneous pinnal injections of antibodies to neutralize PDGF-AB (10 g in 20 L PBS, AB-20-NA, R&D Systems, n 8 cardiac, 8 pulmonary allografts), PDGFR- , PDGFR- (10 g in 20 L PBS, AB-307-NA and AB385, R&D Systems, respectively; n 7 cardiac allografts), or nonimmune rabbit immunoglobulin G (10 g in 20 L PBS, AB-105-C, R&D Systems; n 8 cardiac, 8 pulmonary allografts).

Techniques: Staining, Ligation, Control

Journal: Circulation

Article Title: Platelet-derived growth factor-AB limits the extent of myocardial infarction in a rat model: feasibility of restoring impaired angiogenic capacity in the aging heart.

doi: 10.1161/hc0502.103672

Figure Lengend Snippet: Figure 5. Representative Masson’s trichrome staining in 24-month-old rat hearts pretreated with PBS or PDGF-AB 24 h before LAD ligation. Graph shows myocardial infarct size 14 days after coronary ligation (control, n5; PDGF-AB, n7). *P0.03, PDGF vs control.

Article Snippet: In addition, at the time of cardiac or pulmonary allograft transplantation, sets of young adult mice also were treated with single subcutaneous pinnal injections of antibodies to neutralize PDGF-AB (10 g in 20 L PBS, AB-20-NA, R&D Systems, n 8 cardiac, 8 pulmonary allografts), PDGFR- , PDGFR- (10 g in 20 L PBS, AB-307-NA and AB385, R&D Systems, respectively; n 7 cardiac allografts), or nonimmune rabbit immunoglobulin G (10 g in 20 L PBS, AB-105-C, R&D Systems; n 8 cardiac, 8 pulmonary allografts).

Techniques: Staining, Ligation, Control

Journal: bioRxiv

Article Title: A complex osteoporotic milieu is associated with arterial stiffening and PDGF-BB-mediated calcification of human smooth muscle cells

doi: 10.1101/2025.10.25.684499

Figure Lengend Snippet: Flow cytometry analysis of (A) total granulocytes, (B) basophils, (C) eosinophils, (D) neutrophils, and platelets in the peripheral blood of non-osteoporotic frail controls (CON-F; n=16), osteopenic frail individuals (OPE; n=16), and osteoporotic frail individuals (OPO; n=28 out of 38). Differences in immune cell subsets were assessed using a Brown-Forsythe and Welch ANOVA with Dunnett’s T3 multiple comparisons test vs. CON-F if data were normally distributed. Non-normal data were analyzed with a Kruskal-Wallis test and Dunn’s multiple comparison. § indicates incomplete immune cell subset data. (E) Sera screening of non-osteoporotic middle-aged controls (CON-M), non-osteoporotic healthy older controls (CON-H), CON-F, OPE, and OPO individuals using bead-based multiplex assays. Clustered expression levels are given as median Z-scores. Factors with significantly different expression levels or distinct trends with p<0.1 in OPE or OPO individuals vs. CON-M, CON-H, and CON-F include (G) sCD40L, (H) PDGF-BB, (I) OPN, and (J) SDF-1. Differences between serum factors were assessed using two-way ANOVAs with Dunnett’s post-test (p<0.05) vs. CON-H, CON-M, or CON-F. Only significant differences (p<0.05) and distinct trends with p<0.1 are shown. (K) Spearman correlation of the significantly different cellular and serum factors in non-osteoporotic controls (CON: CON-M, CON-H, CON-F) and (L) in osteopenic and osteoporotic individuals (low BMD: OPE, OPO). (M) Differences between correlation coefficients of CON and low BMD were assessed using Fisher’s Z-test; * p<0.05, ** p<0.001, *** p<0.0001.

Article Snippet: The role of PDGF-BB was further assessed using osteogenic medium containing sex-mixed participant-derived sera pools supplemented with either 25 μg/ml neutralizing polyclonal human PDGF antibody (αPDGF; Cat# AB-20, RRID:AB_354273; R&D Systems ® ) dissolved in PBS, or 50 μM chebulinic acid (CheA; Cat# SMB00271, Sigma-Aldrich or Cat# A16208, AdooQ ® BioScience, USA), dissolved in ddH 2 O and briefly heated up to 70 °C to prepare a 1 mM stock.

Techniques: Flow Cytometry, Comparison, Multiplex Assay, Expressing

Journal: bioRxiv

Article Title: A complex osteoporotic milieu is associated with arterial stiffening and PDGF-BB-mediated calcification of human smooth muscle cells

doi: 10.1101/2025.10.25.684499

Figure Lengend Snippet: SMC undergoing osteogenic differentiation (OM - osteogenic medium) in an in vitro model of SMC calcification were stimulated with recombinant PDGF-BB, sCD40L, OPN, or SDF-1α. Deposited calcified matrix was stained with alizarin red. (A ) Representative images of the deposited calcified matrix. (B-E) Quantified staining, normalized to cell count. A paired, two-tailed t-test was used to assess significant differences (p<0.05) vs. OM without supplements for each time point. (F) Phosphate levels in the supernatant during osteogenic differentiation with the respective recombinant factors. Statistical analysis at each time point was performed using a mixed-effects model with Geisser-Greenhouse correction and Dunnett’s post-test vs. OM without supplements (p<0.05); no significant differences were detected. (G) SMC proliferation in expansion medium (EM; dashed line) after stimulation with the respective recombinant factors. Differences in proliferation were assessed at each time point using two-way ANOVA with Geisser-Greenhouse correction and Dunnett’s post-test vs. EM without supplements (p<0.05); no significant differences were detected. (H) Gene expression analysis of SMC after four days of osteogenic differentiation with the respective recombinant factors. Paired, two-tailed t-tests vs. OM without supplements were used to assess significant differences (p<0.05) across independent experiments.

Article Snippet: The role of PDGF-BB was further assessed using osteogenic medium containing sex-mixed participant-derived sera pools supplemented with either 25 μg/ml neutralizing polyclonal human PDGF antibody (αPDGF; Cat# AB-20, RRID:AB_354273; R&D Systems ® ) dissolved in PBS, or 50 μM chebulinic acid (CheA; Cat# SMB00271, Sigma-Aldrich or Cat# A16208, AdooQ ® BioScience, USA), dissolved in ddH 2 O and briefly heated up to 70 °C to prepare a 1 mM stock.

Techniques: In Vitro, Recombinant, Staining, Cell Counting, Two Tailed Test, Gene Expression

Journal: bioRxiv

Article Title: A complex osteoporotic milieu is associated with arterial stiffening and PDGF-BB-mediated calcification of human smooth muscle cells

doi: 10.1101/2025.10.25.684499

Figure Lengend Snippet: (A) SMC undergoing osteogenic differentiation were stimulated with sex-mixed sera pools of non-osteoporotic healthy older controls (CON-H) or osteoporotic frail individuals with (OPO + ) or without confounders of vascular calcification (OPO - ). PDGF-BB or its downstream signaling was inhibited in the respective sera pools using either 25 µg/ml neutralizing polyclonal PDGF antibody (αPDGF) or 50 µM chebulinic acid (CheA). Deposited calcified matrix was stained with alizarin red, quantified, and normalized to the cell count. A one-way ANOVA with Geisser-Greenhouse correction and Dunnett’s post-test was used to assess significant differences vs. CON-H and the respective osteoporotic group without inhibitor for each time point. (B) Representative images of deposited calcified matrix stained with alizarin red. (C) Phosphate levels determined in the supernatant during osteogenic differentiation. A two-way ANOVA with Geisser-Greenhouse correction and Dunnett’s post-test was used to assess significant differences vs. CON-H for each time point. (D) Cell counts determined by Hoechst staining after stimulation with the respective sera pools supplemented with αPDGF or CheA. A one-way ANOVA with Geisser-Greenhouse correction and Dunnett’s post-test was used to assess significant differences vs. CON-H and the respective osteoporotic group without inhibitor for each time point. Only significant differences (p<0.05) and distinct trends with p<0.1 are shown.

Article Snippet: The role of PDGF-BB was further assessed using osteogenic medium containing sex-mixed participant-derived sera pools supplemented with either 25 μg/ml neutralizing polyclonal human PDGF antibody (αPDGF; Cat# AB-20, RRID:AB_354273; R&D Systems ® ) dissolved in PBS, or 50 μM chebulinic acid (CheA; Cat# SMB00271, Sigma-Aldrich or Cat# A16208, AdooQ ® BioScience, USA), dissolved in ddH 2 O and briefly heated up to 70 °C to prepare a 1 mM stock.

Techniques: Staining, Cell Counting