Journal: Frontiers in pharmacology

Article Title: M2 Macrophages Promote PDGFRβ + Pericytes Migration After Spinal Cord Injury in Mice via PDGFB/PDGFRβ Pathway.

doi: 10.3389/fphar.2021.670813

Figure Lengend Snippet: FIGURE 6 | The expression of PDGFB is significantly increased after SCI or macrophages M2 polarization. (A) Western blot was used to detect the expression levels of PDGFB, PDGFD and PDGFRβ before (Pre) and at 3, 7, and 14 dpi in the injury core of mice (n 5 per group). (B) Quantitative analysis of PDGFB in (A). The blots were quantified as previously described. **p < 0.01, *p < 0.05 (3 and 7 dpi vs. pre); ###p < 0.001, ##p < 0.01 (3 and 7 dpi vs. 14 dpi). (C) Quantitative analysis of PDGFD in (A). ****p < 0.0001 (3, 7, and 14 dpi vs. pre). (D) Quantitative analysis of PDGFRβ in (A). ***p < 0.001 (7, 14 dpi vs. pre and 3 dpi). (E) Western blot was used to detect the expression levels of PDGFB in M0, M1 or M2 macrophages (n 5 per group). (F) Quantitative analysis of PDGFB in (E), the blots were quantified as previously described, **p < 0.01 (M2 vs. M0), ###p < 0.001 (M2 vs. M1). (G) ELISA was used to detect the concentration of PDGFB in CM0, CM1 or CM2 (n 5 per group), The results were expressed as mean ± SD. **p < 0.01 (CM2 vs. other groups).

Article Snippet: According to the experimental requirements, macrophages conditioned medium or PDGFB (10 ng/ml, NBP2-53416, Novus, United States) were added to the lower chamber to regulate MBVP cells migration, or SU16f (10 μm, 3304, R&D Systems, United States) was added to the upper chamber to block PDGFRβ.

Techniques: Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Frontiers in pharmacology

Article Title: M2 Macrophages Promote PDGFRβ + Pericytes Migration After Spinal Cord Injury in Mice via PDGFB/PDGFRβ Pathway.

doi: 10.3389/fphar.2021.670813

Figure Lengend Snippet: FIGURE 7 | M2 macrophages secrete PDGFB acting on PDGFRβ to promote PDGFRβ+ pericytes migration in vitro. (A) Scratch test was used to detect the migration of PDGFRβ+ pericytes after being treated with DMEM (Sham), conditioned medium of M2 macrophages (CM2), 10 ng/ml recombinant mouse PDGFB protein (PDGFB), CM2 plus 10 μm PDGFRβ inhibitor (CM2+SU16f) or PDGFB plus 10 μM PDGFRβ inhibitor (PDGFB+SU16f) for 72 h (n 3 per group). (B) Transwell test was used to further detect the migration of PDGFRβ+ pericytes after being treated in Sham, CM2, PDGFB, CM2 + SU16f or PDGFB+SU16f groups for 20 h (n 3 per group). (C) Quantitative analysis of the wound closure rate in (A), the results were quantified as previously described, ****p < 0.0001, ***p < 0.001 (groups vs. Sham), ####p < 0.0001 (groups vs. CM2 and PDGFB). (D) Quantitative analysis of the number of transmembrane cells in (B), the results were quantified as previously described, ****p < 0.0001 (CM2 and PDGFB vs. other groups). ND, not determined. Scale bars, 100 μm. (E) Schematic representation. M2 macrophages could secrete PDGFB, acting on PDGFRβ of PDGFRβ+ pericytes, which promote the formation of fibrotic scar, corral macrophages and limit inflammation after SCI.

Article Snippet: According to the experimental requirements, macrophages conditioned medium or PDGFB (10 ng/ml, NBP2-53416, Novus, United States) were added to the lower chamber to regulate MBVP cells migration, or SU16f (10 μm, 3304, R&D Systems, United States) was added to the upper chamber to block PDGFRβ.

Techniques: Migration, In Vitro, Recombinant

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Regulation of seminiferous tubule-associated stem Leydig cells in adult rat testes

doi: 10.1073/pnas.1519395113

Figure Lengend Snippet: Factors screened by tubule culture experiments for their roles in the proliferation and differentiation of stem Leydig cells

Article Snippet: This same sequence occurs in vivo ( 21 , 22 ). table ft1 table-wrap mode="anchored" t5 Table S1. caption a7 Reagent Signaling pathway Concentration (range tested) Manufacturer Ovine LH 10 ng/mL (0.1–10) Hormone Program USDA Rat FSH 20 ng/mL (0.2–20) Hormone Program NIDDK T3 10 nM (0.1–10) Sigma-Aldrich Estradiol (E2) 10 ng/mL (0.1–10) Steraloids DHT 10 ng/mL (0.1–10) Steraloids R1881 AR − 10 nM (0.1–10) Sigma-Aldrich Dexamethasone 10 nM (0.1–10) Sigma-Aldrich Retinoic acid 1 μM (0.01–1) Sigma-Aldrich Mouse PDGFAA 10 ng/mL (0.1–10) Prospec Protein Specialists Rat PDGFBB 10 ng/mL (0.1–10) Sigma-Aldrich PDGFR Inhibitor IV PDGFBB − 20 nM (0.2–20) EMD Bioscience Imatinib Mesylate c-Kit − /PDGFR − 0.2 μM (0.02–2) Selleck Chemicals Human TGFβ1 10 ng/mL (0.1–10) Cell Signaling Technology Human Activin 10 ng/mL (0.1–10) Sigma-Aldrich SB431542 TGFβ − /Activin − 0.1 μM (0.01–1) Selleck Chemicals SB525334 TGF-β − 50 nM (0.5–50) Selleck Chemicals Rat TGFα 10 ng/mL (0.1–10) Sigma-Aldrich Mouse EGF 10 ng/mL (0.1–10) Sigma-Aldrich Porcine Inhibin 10 ng/mL (0.1–10) Sigma-Aldrich Rat IL-1β 1 ng/mL (0.1–10) Sigma-Aldrich Rat IGF1 50 ng/mL (0.5–50) Sigma-Aldrich Human SCF 10 ng/mL (0.1–10) MP Biomedicals Rat GDNF 10 ng/mL (0.1–10) EMD Chemicals Human FGF2 10 ng/mL (0.1–10) Novus Biologiclas Mouse LIF 10 ng/mL (0.1–10) Sigma-Aldrich DAPT Notch − 2 μM (0.2–20) EMD Bioscience Lithium ?

Techniques: