Journal: Cell Communication and Signaling : CCS

Article Title: GDF9 His209GlnfsTer6/S428T and GDF9 Q321X/S428T bi-allelic variants caused female subfertility with defective follicle enlargement

doi: 10.1186/s12964-024-01616-8

Figure Lengend Snippet: Effects of GDF9 variants on gene expression and GDF9-BMP15 interactions. A Western blots of mature GDF9 proteins expressed in follicular fluid collected from the first and second retrieval cycle of individual P1 and age-matched controls ( n =3). B ELISA of GDF9 in follicular fluid collected from the first retrieval cycle of individual P1 and age-matched controls ( n =6). C Western blots of GDF9 expression in HEK293T cells transfected with human GDF9 WT , GDF9 Q321X , GDF9 S428T , and GDF9 His209GlnfsTer6 respectively and their relative mature-GDF9 expression in culture medium (CM) (3 independent experiments). The amount of concentrated culture medium used for the detection was 35μl per sample per assay. lnfsTer6, His209GlnfsTer6. * P < 0.05. D Western blots of GDF9 precursors and mature GDF9 over time during in vitro cleavage assay (3 independent experiments). Total GDF9=GDF9 precursor+mature GDF9. E Western blots of BMP15 expression in HEK293T cells co-transfected with human BMP15 and human GDF9 WT , GDF9 Q321X , GDF9 S428T , and GDF9 His209GlnfsTer6 respectively and their relative BMP15 precursor expression in CM (3 independent experiments). lnfsTer6, His209GlnfsTer6. * P < 0.05. F Co-IP assay of the binding of GDF9 proteins to BMP15 protein. lnfsTer6, His209GlnfsTer6. * P < 0.05. G Western blots of p-smad2 and smad2 expression in human primary luteinized granulosa cells treated with purified recombinant pro-GDF9 WT -BMP15 and pro-GDF9 S428T -BMP15 protein respectively (3 independent experiments)

Article Snippet: GDF9 was measured in follicular fluid collected at the first oocyte retrieval from individual P1 and in follicular fluid from other six control women using an ELISA kit (CSB-E12925h, CUSABIO, China).

Techniques: Gene Expression, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Transfection, In Vitro, Cleavage Assay, Co-Immunoprecipitation Assay, Binding Assay, Purification, Recombinant

Journal: Journal of Tissue Engineering

Article Title: Therapeutic potential and mechanisms of umbilical cord mesenchymal stem cells differentiating into tendon cells and promotion of rotator cuff tendon-bone healing

doi: 10.1177/20417314251315185

Figure Lengend Snippet: Analysis of the induction effect of BMP-12, GDF-7, and CTGF on the differentiation of UCMSCs: (a) morphological changes of UCMSCs after induction treatment, (b) mRNA expression of MKX, SCX, and TNC detected by qPCR, (c) bands and protein expression of MKX, SCX, and TNC detected by WB, and (d) COL-I and COL-III expression detected by immunofluorescence (400×). ** p < 0.01 and *** p < 0.001 compared with the control group.

Article Snippet: Rat umbilical cord mesenchymal stem cells (UCMSCs, CP-R302, Pricella, China), 293T cells (Pricella, China), DMEM medium (L110KJ, Yuanpei, China), Fetal bovine serum(AC03L055, life-ilab, China), GDF-6 (HY-P79333, MedChemexpress, USA), GDF-7 (92004ES10, Yisheng, China), CTGF (HY-P72154, MedChemexpress Biotechnology, USA), GoldenstarTM RT6 cDNA Synthesis Kit Ver.2 (TSK302M, Tsingke, China), qPCR Mix (SYBR Green I) kit and PCR primer (TSE002, Tsingke, China), RIPA lysis buffer (Beyotime, China), TNC (A1927, abclonal, China), MKX (ab236400, abcam, UK), SCX (ab58655, abcam, UK), GAPDH (A19056, abcam, UK), Hes1 (ab71559, abcam, UK), Hras (ab32417, abcam, UK), GAPDH Rabbit mAb (A19056, abclonal, China), Lentiviral Packaging Kit (41102ES10, Yisheng, China), Fetal bovine serum (26050070, Thermo, USA), trypsin (LP0042, Thermo, USA), PBS (10010001, Thermo, USA).

Techniques: Expressing, Immunofluorescence, Control

Journal: Journal of Cachexia, Sarcopenia and Muscle

Article Title: Fibro‐Adipogenic Progenitors Regulate Orofacial Neuromuscular Junction Regeneration via Myostatin

doi: 10.1002/jcsm.70264

Figure Lengend Snippet: Recombinant MSTN suppressed AChR clustering and miR‐206 expression. (a) AChR and MyHC staining of MuSCs cultures treated with or without recombinant MSTN (100 ng/mL) and follistatin (500 ng/mL). Scale bar = 50μm. (b) Graphs show (from left to right) the quantification of differentiation index, myotube area, AChR cluster counts normalized to myotube area and ratio of AChR/myotube area. n = 3. (c) The levels of miR‐206 expression in MuSCs cultures. Mature myotubes differentiated from MAS‐derived MuSCs were treated with control, agrin alone, agrin plus MSTN or agrin plus MSTN and follistatin. (d) AChR and MyHC staining of myotubes transfected with miR‐206 mimics or negative control (NC) prior to treatment with agrin and MSTN. Scale bar = 50 μm. (e) Graphs show (from left to right) the quantification of differentiation index, myotube area, AChR cluster counts normalized to myotube area and ratio of AChR/myotube area. n = 3. The data are shown as mean ± SD. One‐way ANOVA followed by Tukey's post hoc test was used. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Mature myotubes were treated with 100 ng/mL agrin (HY‐ P79236 , MedChemExpress, China) in DM or conditioned medium (CM) of 7 dpi FAPs supplemented with 2% HS for 16 h. For recombinant MSTN treatment, 100 ng/mL MSTN (HY‐ P72632 , MedChemExpress, China) with or without 500 ng/mL follistatin (HY‐ P70315 , MedChemExpress, China) was administered together with agrin treatment.

Techniques: Recombinant, Expressing, Staining, Derivative Assay, Control, Transfection, Negative Control

Journal: bioRxiv

Article Title: Therapy-induced senescent-like cancer cells drive macrophage-mediated immunosuppression in cholangiocarcinoma

doi: 10.64898/2026.06.24.734341

Figure Lengend Snippet: (A) Proteome Profiler Human XL cytokine array (105 cytokines) of conditioned medium from proliferating, doxorubicin-induced (Dox), and Gem/Cis-induced senescent-like RBE cells. (B) Gdf15 mRNA in proliferating versus senescent-like murine CCA (SB1) cells. (C) Gdf15 mRNA in control vs AP-treated SB-p16-ATTAC tumors. (D) Schematic of in vitro studies. (E-F) Arg1 and iNOS mRNA in bone-marrow-derived macrophage (BMDM) incubated for 72 h with conditioned medium from vehicle- or alisertib-treated SB1 cells expressing shCtrl or shGdf15 . (G) Bulk RNA-seq heatmap (row Z-scores) of naïve bone marrow-derived macrophages (BMDM) cultured 72 h with conditioned media (CM) from SB-p16-shCtrl or SB- p16-Gdf15 OE cells. (H) ARG1 mRNA in human monocyte-derived macrophages ± recombinant human GDF-15 (Qkine Qk017; TGF-β1-free), qRT-PCR relative to vehicle; mean ± SEM, n = 6. (I) Division index (CFSE dilution, normalized to T-cells-only) of anti-CD3/CD28-activated CD8⁺ T cells co-cultured with BMDMs incubated for 72 h with conditioned medium from vehicle- or alisertib-treated SB1 cells expressing shCtrl or shGdf15 . Data are mean ± SEM; *, P<0.05; **, P<0.01; ****, P<0.0001; ns, not significant.

Article Snippet: Human recombinant GDF-15 was from Qkine (Qk017) and murine recombinant GDF-15 from R&D Systems (8944-GD/CF).

Techniques: Control, In Vitro, Derivative Assay, Incubation, Expressing, RNA Sequencing, Cell Culture, Recombinant, Quantitative RT-PCR

Journal: bioRxiv

Article Title: Therapy-induced senescent-like cancer cells drive macrophage-mediated immunosuppression in cholangiocarcinoma

doi: 10.64898/2026.06.24.734341

Figure Lengend Snippet: (A) Macrophages express TGFBR2, the candidate non-canonical GDF-15 receptor, in murine and human CCA single-cell RNA-seq. (B) α-TGFBR2 western blot (WB) of streptavidin pull-down (top) and input (middle) from naïve BMDMs treated with biotin-supplemented conditioned media (CM) from CCA cells expressing GDF-15-BioID2-HA or SEC61B-BioID2-HA (ER control); α-HA blot of CM (bottom) confirms bait secretion. (C) GDF-15 induces macrophage phospho-STAT6 but not phospho-SMAD2/3, reflecting non-canonical TGFBR2 output. (D) WB of BMDMs stimulated with control or GDF-15-overexpressing (GDF-15 OE ) CM for 0 or 1 h: p-STAT6, STAT6, p-SMAD3, SMAD3. (E) Arg1 mRNA in BMDMs transfected with siCtrl or siTgfbr2 and stimulated with control or rGDF-15. n = 6. Data are mean ± SEM; *, P<0.05; **, P<0.01; ****, P<0.0001; ns, not significant.

Article Snippet: Human recombinant GDF-15 was from Qkine (Qk017) and murine recombinant GDF-15 from R&D Systems (8944-GD/CF).

Techniques: Single Cell, RNA Sequencing, Western Blot, Expressing, Control, Transfection

Journal: bioRxiv

Article Title: Therapy-induced senescent-like cancer cells drive macrophage-mediated immunosuppression in cholangiocarcinoma

doi: 10.64898/2026.06.24.734341

Figure Lengend Snippet: (A) Strategy for p16-promoter-restricted Gdf15 silencing (shRNA) and Gdf15 overexpression. (B) qPCR validation of Gdf15 knockdown and overexpression in senescent-like SB1 cells. (C-D) Subcutaneous model and tumor-burden quantification. (E) GeoMx spatial profiling of GDF-15 expression and macrophage infiltration in human CCA. Region-of-interest (ROI)-level analysis plotting GDF-15 expression against macrophage score; each point is one ROI, with the linear fit and 95% confidence band shown. Spearman ρ = 0.63, P = 0.0067. Data are mean ± SEM; **, P<0.01; ****, P<0.0001.

Article Snippet: Human recombinant GDF-15 was from Qkine (Qk017) and murine recombinant GDF-15 from R&D Systems (8944-GD/CF).

Techniques: shRNA, Over Expression, Biomarker Discovery, Knockdown, Expressing

Journal: Stem Cell Research & Therapy

Article Title: Pretreatment with growth differentiation factor 15 augments cardioprotection by mesenchymal stem cells in myocardial infarction by improving their survival

doi: 10.1186/s13287-024-04030-6

Figure Lengend Snippet: Antibody information

Article Snippet: For GDF15 (MedChemExpress, HY-P75170) pretreatment, MSCs were cultured on fresh completed medium supplemented with GDF15 (10, 20, 50 and 100 ng/mL) under normoxia (95% air and 5% CO 2 ) for 24 h and then exposed to serum deprivation and hypoxia (SD/H, 1% oxygen, 5% carbon dioxide and 94% nitrogen) for 48 h to mimic the ischemic conditions in vitro.

Techniques:

Journal: Stem Cell Research & Therapy

Article Title: Pretreatment with growth differentiation factor 15 augments cardioprotection by mesenchymal stem cells in myocardial infarction by improving their survival

doi: 10.1186/s13287-024-04030-6

Figure Lengend Snippet: GDF15 pretreatment augmented paracrine effects of MSCs. A Schematic showing the experiments for assaying the paracrine effect of MSCs with or without GDF15. B Representative images of tube formation in HUVECs treated with DMEM, MSC-CM or GDF15-MSC-CM. Scale bar = 100 μm. C Quantification of tube formation in HUVECs treated with DMEM, MSC-CM or GDF15-MSC-CM. D The concentration of VEGF, EGF and bFGF in MSC-CM and GDF15-MSC-CM was determined by ELISA assay. E Representative images of the apoptotic NCMs treated with DMEM, MSC-CM or GDF15-MSC-CM under SD/H condition. Scale bar = 200 μm. F Quantification of the apoptotic rate of NCMs treated with DMEM, MSC-CM or GDF15-MSC-CM under SD/H condition. Data are expressed as the mean ± SEM. N = 3 biological replicates for each group. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: For GDF15 (MedChemExpress, HY-P75170) pretreatment, MSCs were cultured on fresh completed medium supplemented with GDF15 (10, 20, 50 and 100 ng/mL) under normoxia (95% air and 5% CO 2 ) for 24 h and then exposed to serum deprivation and hypoxia (SD/H, 1% oxygen, 5% carbon dioxide and 94% nitrogen) for 48 h to mimic the ischemic conditions in vitro.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Stem Cell Research & Therapy

Article Title: Pretreatment with growth differentiation factor 15 augments cardioprotection by mesenchymal stem cells in myocardial infarction by improving their survival

doi: 10.1186/s13287-024-04030-6

Figure Lengend Snippet: Transplantation of GDF15-MSCs improved cardiac function following infarction in mice. A Representative images of echocardiography taken at 28 days in Sham or mice with MI that received transplantation of PBS, MSCs, or GDF15-MSCs. B Quantitative analysis of LVEF and LVFS at baseline (before MI) and 28 days later in Sham or mice with MI that received transplantation of PBS, MSCs or GDF15-MSCs. C Representative Masson’s trichrome staining images of hearts of Sham mice or mice with MI that received transplantation of PBS, MSCs or GDF15-MSCs. Scale bar = 2 mm. D Quantitative analysis of infarct size of heart from Sham mice or mice with MI that received transplantation of PBS, MSCs or GDF15-MSCs. E Representative images of anti-human mitochondria staining of heart tissue from mice with MI that received PBS, MSCs or GDF15-MSCs treatment and sham group. Scale bar = 50 μm. F Quantitative analysis of MSC survival in heart tissue of mice with MI that received transplantation of MSCs or GDF15-MSCs. Data are expressed as the mean ± SEM. N = 6. *p < 0.05; **p < 0.01; ***p < 0.001

Article Snippet: For GDF15 (MedChemExpress, HY-P75170) pretreatment, MSCs were cultured on fresh completed medium supplemented with GDF15 (10, 20, 50 and 100 ng/mL) under normoxia (95% air and 5% CO 2 ) for 24 h and then exposed to serum deprivation and hypoxia (SD/H, 1% oxygen, 5% carbon dioxide and 94% nitrogen) for 48 h to mimic the ischemic conditions in vitro.

Techniques: Transplantation Assay, Staining

Journal: Stem Cell Research & Therapy

Article Title: Pretreatment with growth differentiation factor 15 augments cardioprotection by mesenchymal stem cells in myocardial infarction by improving their survival

doi: 10.1186/s13287-024-04030-6

Figure Lengend Snippet: Transplantation of GDF15-MSCs inhibited cardiomyocyte apoptosis and enhanced angiogenesis in a mouse model of MI. A Representative images of TUNEL and Troponin double staining of heart tissue from Sham mice or mice with MI that received transplantation of PBS, MSCs, or GDF15-MSCs. Scale bar = 100 μm. B Quantitative analysis of cardiomyocyte apoptosis in heart tissue from the different groups. C Representative images of CD45 staining of heart tissue from Sham mice or mice with MI that received transplantation of PBS, MSCs, or GDF15-MSCs. Scale bar = 100 μm. D Quantitative analysis of CD45 density in heart tissue from the different groups. E Representative images of CD31 and α-SMA staining of heart tissue from Sham mice or mice with MI that received transplantation of PBS, MSCs or GDF15-MSCs. Scale bar = 200 μm. F Quantitative analysis of CD31 density in heart tissue from the different groups. G Quantitative analysis of α-SMA density in heart tissue from the different groups. Data are expressed as the mean ± SEM. N = 6. **p < 0.01; ***p < 0.001

Article Snippet: For GDF15 (MedChemExpress, HY-P75170) pretreatment, MSCs were cultured on fresh completed medium supplemented with GDF15 (10, 20, 50 and 100 ng/mL) under normoxia (95% air and 5% CO 2 ) for 24 h and then exposed to serum deprivation and hypoxia (SD/H, 1% oxygen, 5% carbon dioxide and 94% nitrogen) for 48 h to mimic the ischemic conditions in vitro.

Techniques: Transplantation Assay, TUNEL Assay, Double Staining, Staining