su 16f  (MedChemExpress)

Journal: bioRxiv

Article Title: Suppressing PDGFRβ Signaling Enhances Myocyte Fusion to Promote Skeletal Muscle Regeneration

doi: 10.1101/2024.10.15.618247

Figure Lengend Snippet: (A) Schematic overview of the experimental procedure. Hindlimb muscles were collected from P30 male mice, and isolated muscle progenitor cells (MPCs) were cultured. Throughout differentiation, cells were treated with vehicle, PDGF-BB (25 ng/mL), or SU16f (1 µM) for five days and then myotube formation was assessed. (B) Representative images of myosin heavy chain (MyHC) showing myotube development in cultures as described in (A), providing visual evidence of the effects of PDGF-BB and SU16f on myotube formation. (C) Quantification of the fusion index of differentiated cultures described in (B), reflecting the efficiency of myoblast fusion into myotubes in the treated cultures (n = 3 biologically independent mice per group). (D) Quantification of myotube nuclei count and distribution from cultures described in (B) (n = 4 biologically independent mice per group). (E) Muscle progenitor cells were isolated from hindlimb muscles of MyoG tdTomato mice and cultured at low density. Cells were treated with vehicle, PDGF-BB (25 ng/mL), or SU16f (1 µM), and the differentiation index was determined by monitoring MyoG tdTomato positivity (n = 4 biologically independent mice per group). (F) Schematic overview of the regenerative experimental procedure. MyoG tdTomato mice were subjected to a single intramuscular injection of 1.2% BaCl₂ to induce injury in the TA muscle. Subsequently, mice were administered one dose of vehicle, PDGF-BB (50 ng/mouse), or SU16f (2 mg/Kg) for five consecutive days by intraperitoneal injection. Muscle regeneration was assessed at seven d.p.i. (G) Representative images of TA muscle sections stained for eMyHC and laminin, showing the extent of muscle regeneration and myofiber repair in the different experimental conditions described in (F). (H) Quantification of injured myofiber cross sectional area (CSA) from the images described in (G), providing a comparative analysis of muscle regeneration efficiency (n = 4 biologically independent mice per group). (I) Quantification of myonuclear accretion per injured myofiber, offering insights into the cellular response to injury and subsequent repair (n = 4 biologically independent mice per group). (J) Quantification of eMyHC immunostaining from muscle sections described in (G), indicating the level of ongoing myofiber regeneration in the different experimental conditions (n = 4 biologically independent mice per group). Data are presented as mean ± SEM, with each experimental group consisting of biologically independent replicates as indicated. Statistical significance was determined using a one-way ANOVA for panels (C, E, H, and J) or a two-way ANOVA for panels (D) and (I). Scale bar = 100 µm.

Article Snippet: For the denoted experiments, cells were treated with either vehicle (0.1% DMSO), 25 ng Pdgfrβ ligand (PDGF-BB; VWR 10780-774), 1 µM SU16F (Tocris 3304), or 1 µM Fludarabine (Tocris Bioscience: 3495).

Techniques: Muscles, Isolation, Cell Culture, Injection, Staining, Immunostaining

Journal: bioRxiv

Article Title: Suppressing PDGFRβ Signaling Enhances Myocyte Fusion to Promote Skeletal Muscle Regeneration

doi: 10.1101/2024.10.15.618247

Figure Lengend Snippet: (A) Schematic of the experimental approach. Muscle progenitor cells were FACS-isolated from human quadricep muscles and cultured. These cells were treated with vehicle, PDGF-BB (25 ng/mL), or SU16f (1 µM) throughout the myogenesis process. Myotube development was assessed to evaluate the effects of these treatments. (B) Representative images of myotube development as assessed by MyHC immunostaining from human muscle cell cultures described in (A). (C) Quantification of the fusion index from cultures and images described in (A and B) offering insights into human muscle cell fusion under different treatment conditions (n = 5 biologically independent samples per group). (D) Quantification of nuclei and distribution within myotubes from images described in (B), reflecting myonuclear accretion in response to targeting PDGFRβ (n = 5 biologically independent samples per group). (E) Quantification of myotube length from cultures described in (A) and (B), providing a measure of myotube development under the influence of PDGF-BB and SU16f (n = 5 biologically independent samples per group). (F) Quantification of myotube diameter from the cultures described in (A) and (B), indicating the effect of the treatments on the thickness of the developing myotubes (n = 5 biologically independent samples per group). Data are presented as mean values with individual data points ± S.E.M. Statistical significance was determined using a one-way ANOVA for panels (C-F). Scale bar = 100 µm.

Article Snippet: For the denoted experiments, cells were treated with either vehicle (0.1% DMSO), 25 ng Pdgfrβ ligand (PDGF-BB; VWR 10780-774), 1 µM SU16F (Tocris 3304), or 1 µM Fludarabine (Tocris Bioscience: 3495).

Techniques: Isolation, Muscles, Cell Culture, Immunostaining

Journal: Research

Article Title: PDGFR in PDGF-BB/PDGFR Signaling Pathway Does Orchestrates Osteogenesis in a Temporal Manner

doi: 10.34133/research.0086

Figure Lengend Snippet: PDGF-BB activates the inhibitory effect of Erk 1/2 on the pathway of Smad-1/5/9 and promotes cell proliferation activity rather than osteogenic characterization while the PDGFR-β inhibitor (SU-16f) has the opposite effect. (A) ALP staining of the groups (control, black; 10 ng/ml PDGF-BB, red; 5 μM PDGFR-β inhibitor, purple) with low (40×) and high (100×) magnification of optical micrograph. Scale bar, 50 μm. (B) ARS of the groups (control, black; 10 ng/ml PDGF-BB, red; 5 μM PDGFR-β inhibitor, purple) with low (40×) and high (100×) magnification of optical micrograph. Scale bar, 50 μm. (C) Quantitative analysis of ALP staining of the groups. Data are means ± SEM. One-way ANOVA. *** P < 0.001. (D) Quantitative analysis of ARS of the groups. Data are means ± SEM. One-way ANOVA. ** P < 0.01; *** P < 0.001. (E) EdU assay of the groups. (F) EdU assay of the groups in different time points (1, 3, and 5 d) with time trend and quantitative analysis. Data are means ± SEM. One-way ANOVA. ** P < 0.01; *** P < 0.001. (G) PCR analysis of ALP, Col1a1, and Runx-2 mRNA of the groups with 5 d. Data are means ± SEM. One-way ANOVA. ns P ≥ 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001. (H) PCR analysis of ALP, Col1a1, and Runx-2 mRNA of the groups with 7 d. Data are means ± SEM. One-way ANOVA. ns P ≥ 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001. (I) WB band of Smad 1/5/9 and Smad 1 with 7 d.

Article Snippet: PDGF-BB was purchased from R&D Systems (Shanghai, China), and PDGFR-β inhibitor, also called SU-16f, was purchased from MedChemExpress (USA).

Techniques: Activity Assay, Staining, Control, EdU Assay

Journal: Research

Article Title: PDGFR in PDGF-BB/PDGFR Signaling Pathway Does Orchestrates Osteogenesis in a Temporal Manner

doi: 10.34133/research.0086

Figure Lengend Snippet: Blocked PDGFR-β in the late stage of osteogenesis can promote osteogenic function of hMSCs. (A) The schematic diagram of temporal control grouping with total time of 8 d (number before P or S respectively means cultured days of 10 ng/ml PDGF-BB or 5 μM SU-16f in the medium). (B) ALP staining with low (40×) and high (100×) magnification of optical micrograph. Scale bar, 50 μm. (C) Quantitative analysis of ALP analysis (control, black; 0P-8S, lilac; 2P-6S, modena; 4P-4S, amaranth; 6P-2S, light red; 8P-0S, red). Data are means ± SEM. n = 5 or 6. One-way ANOVA. ns P ≥ 0.05; ** P < 0.01. (D) The schematic diagram of temporal control grouping with total time of 12 d (number before P or S respectively means cultured days of 10 ng/ml PDGF-BB or 5 μM SU-16f in the medium). (E) ARS with low (40×) and high (100×) magnification of optical micrograph. Scale bar, 50 μm. (F) Quantitative analysis of ARS (control, black; 0P-12S, lilac; 3P-9S, modena; 6P-6S, amaranth; 9P-3S, light red; 12P-0S, red). Data are means ± SEM. n = 5 or 6. One-way ANOVA. ns P ≥ 0.05; ** P < 0.01. (G) The schematic diagram of vivo experiment grouping (number before P or S respectively means administration weeks of nontreatment or SU-16f). (H) The quantitative analysis of micro-CT reconstruction (control [0P-8S], black, given PBS by intraperitoneal injection [ip.] once every 2 d [qod.] after skull operation; 6P-2S, amaranth, given 10 ng SU-16f by ip. qod. in the 7th and 8th weeks after skull operation; 4P-4S, lilac, given 10 ng SU-16f by ip. qod. from 5th to 8th week after skull operation; 2P-6S, light red, given 10 ng SU-16f by ip. qod. from 3rd to 8th week after skull operation; 0P-8S, modena, given 10 ng SU-16f by ip. qod. after skull operation). Data are means ± SEM. n = 5 or 6. One-way ANOVA. ns P ≥ 0.05; ** P < 0.01. (I) The reconstructions of representative calvarial models (average of the individual samples) measured by micro-CT. Original defect area is shaded with a dashed outline (control [0P-8S], black dotted line; 6P-2S, amaranth dotted line; 4P-4S, lilac dotted line; 2P-6S, light red dotted line; 0P-8S, modena dotted line).

Article Snippet: PDGF-BB was purchased from R&D Systems (Shanghai, China), and PDGFR-β inhibitor, also called SU-16f, was purchased from MedChemExpress (USA).

Techniques: Control, Cell Culture, Staining, Micro-CT, Injection

Journal: Research

Article Title: PDGFR in PDGF-BB/PDGFR Signaling Pathway Does Orchestrates Osteogenesis in a Temporal Manner

doi: 10.34133/research.0086

Figure Lengend Snippet: The administration of PDGFR-β inhibitor (SU-16f) in the late stage of osteogenesis of bone CSD mouse can significantly promote bone healing and mineralization with assistance of ECM scaffold. (A) The drug delivery schematic diagram of control and PDGFR-β inhibitor serious groups with dose control (Low dose, 10 ng SU-16f qod. ip.; High dose, 100 ng SU-16f qod. ip.) and temporal control (Early, the 1st to 4th week after skull CSD operation; Late, the 5th to 8th week after skull CSD operation). (B) The reconstructions of representative calvarial models (average of the individual samples) measured by micro-CT. Original defect area is shaded with a dashed outline (control, black dotted line, given PBS by ip. qod. after skull operation; Early/Low, purple single dotted line, given 10 ng SU-16f, ip. qod. from the 1st to 4th week; Early/High, purple double dotted line, given 100 ng SU-16f, ip. qod. from the 1st to 4th week; Late/Low, purple single full line, given 10 ng SU-16f ip. qod. from the 5th to 8th week; Late/High, purple double full line, given 100 ng SU-16f ip. qod. from the 5th to 8th week). (C) The quantitative analysis of micro-CT reconstruction (control, black; Early/Low, lilac without board; Early/High, modena without board; Late/Low, lilac with board; Late/High, modena with board line). Data are means ± SEM. n = 5 or 6. One-way ANOVA. ns P ≥ 0.05; ** P < 0.01. (D) The H&E staining image of each group (control, black dotted line; Early/Low, purple single dotted line; Early/High, purple double dotted line; Late/Low, purple single full line; Late/High, purple double full line). Scale bar, 50 μm. (E) The Masson staining of each group (control, black dotted line; Early/Low, purple single dotted line; Early/High, purple double dotted line; Late/Low, purple single full line; Late/High, purple double full line). Scale bar, 50 μm.

Article Snippet: PDGF-BB was purchased from R&D Systems (Shanghai, China), and PDGFR-β inhibitor, also called SU-16f, was purchased from MedChemExpress (USA).

Techniques: Control, Micro-CT, Staining

Journal: Research

Article Title: PDGFR in PDGF-BB/PDGFR Signaling Pathway Does Orchestrates Osteogenesis in a Temporal Manner

doi: 10.34133/research.0086

Figure Lengend Snippet: PDGFR-β inhibitor (SU-16f) which does not rely on biological materials and high dosage can also effectively promote bone healing in mice. (A) The drug delivery schematic diagram of control and PDGFR-β inhibitor with high dose (100 ng SU-16f, qod., ip.) and low dose (10 ng SU-16f, qod., ip.). (B) The reconstructions of representative calvarial models (average of the individual samples) measured by micro-CT. Original defect area is shaded with a dashed outline (control, black single dotted line; Low dose, purple single dotted line; High dose, purple double dotted line). (C) The quantitative analysis of micro-CT reconstruction of CSD model. Data are means ± SEM. n = 5 or 6. One-way ANOVA. ns P ≥ 0.05; ** P < 0.01. (D) The drug delivery schematic diagram of control and PDGFR-β inhibitor (10 ng SU-16f, qod., ip.). (E) The reconstructions of representative calvarial models (average of the individual samples) measured by micro-CT. Original defect area is shaded with a dashed outline (control, black; PDGFR-β inhibitor, purple). (F) The quantitative analysis of micro-CT reconstruction of CSD model. Data are means ± SEM. n = 5 or 6. Student t test. ns P ≥ 0.05; * P < 0.05; ** P < 0.01; *** P < 0.001. (G) The H&E staining image of each group (control, purple dotted line; PDGFR-β inhibitor, purple full line). Scale bar, 50 μm. (H) The Masson staining of each group (control, red dotted line; PDGFR-β inhibitor, red full line). Green triangle arrow, small blood vessels. Scale bar, 50 μm.

Article Snippet: PDGF-BB was purchased from R&D Systems (Shanghai, China), and PDGFR-β inhibitor, also called SU-16f, was purchased from MedChemExpress (USA).

Techniques: Control, Micro-CT, Staining

Journal: Research

Article Title: PDGFR in PDGF-BB/PDGFR Signaling Pathway Does Orchestrates Osteogenesis in a Temporal Manner

doi: 10.34133/research.0086

Figure Lengend Snippet: Immunofluorescence shows that the time-controlled delivery of PDGFR-β inhibitor (SU-16f) does not affect formation of blood vessels in the bone defect but increases osteogenic targets. (A and B) The fluorescence images with CD31 (white) and Dapi (blue) (2 groups with both white full line). Scale bar, 100 μm. (C and D) The fluorescence images with Erk 1/2 (amaranth) and Dapi (blue) (control, full line’PDGFR-β inhibitor, dotted line). (E and F) The fluorescence images with Smad 1/5/9 (green) and Dapi (blue) (control, dotted line; PDGFR-β inhibitor, full line). (G and H) The fluorescence images with Col1a1 (yellow) and Dapi (blue) (control, dotted line; PDGFR-β inhibitor, full line). (I and J) The fluorescence images with Runx-2 (cyan) and Dapi (blue) (control, dotted line, PDGFR-β inhibitor, full line). (K and L) The fluorescence images with Osterix (red) and Dapi (blue) (control, dotted line; PDGFR-β inhibitor, full line).

Article Snippet: PDGF-BB was purchased from R&D Systems (Shanghai, China), and PDGFR-β inhibitor, also called SU-16f, was purchased from MedChemExpress (USA).

Techniques: Immunofluorescence, Fluorescence, Control

Journal: Research

Article Title: PDGFR in PDGF-BB/PDGFR Signaling Pathway Does Orchestrates Osteogenesis in a Temporal Manner

doi: 10.34133/research.0086

Figure Lengend Snippet: The abstract figure of the effect of PDGFR-β inhibitor (SU-16f) on pathways in late stage of bone healing. (A) The early stage of osteogenic repair focuses on cell expansion and angiogenesis, while the later stage should emphasize osteogenic differentiation and mineralization. (B) The abstract figure shows the role of PDGF-BB in bone repair in general. (C) The abstract figure shows SU-16f (PDGFR-β inhibitor) antagonizes PDGFR-β to promote bone repair.

Article Snippet: PDGF-BB was purchased from R&D Systems (Shanghai, China), and PDGFR-β inhibitor, also called SU-16f, was purchased from MedChemExpress (USA).

Techniques: