Journal: Cancers

Article Title: Visfatin Mediates Malignant Behaviors through Adipose-Derived Stem Cells Intermediary in Breast Cancer

doi: 10.3390/cancers12010029

Figure Lengend Snippet: GDF15 play a crucial role in the regulations of breast cancer cells activities by visfatin-treated ADSCs (vADSCs). ( A ) After the three-day co-culture of MDA-MB-231 cells and vADSCs (V50 and V100 group) or untreated ADSCs (Ctrl group), the CM was collected and analyzed by using a cytokine array kit. ( B ) The expression of GDF15 in the co-cultured CM was validated by ELISA represented in a histogram. The ADSCs and MDA-MB-231 cells collected from the co-culture system were extracted for cell lysate to analyze the GDF15 expression by western blotting. ( C ) The migration and invasion of MDA-MB-231 treated with GDF15 at various concentrations for 48 h were evaluated by using a transwell system. ( D ) The indirect co-culture was performed in the presence or absence of the GDF15 neutralizing antibody of for three days. After that, the migration and invasion of the MDA-MB-231 cells collected from the co-culture were evaluated in a transwell system. ( E ) The expression of phosphor-AKT (pAKT) of MDA-MB-231 treated with GDF15 (50 ng/mL) at different time point was detected by western blotting. ( F ) The pAKT was detected in the MDA-MB-231 cells from the co-culture by western blotting. ( G ) After the three-day co-culture in the presence or absence of the wortmannin (400 nM), the MDA-MB-231 cells were collected from the co-culture for performing the migration assay. All experiments were performed in triplicate.

Article Snippet: For performing cell invasion or migration assay, the MDA-MB-231 cells collected from the co-culture system or treated with human recombinant GDF15 (10936-H01H, Sino Biological, Wayne, PA, USA) were suspended in serum-free DMEM and transferred to the 24-well transwell inserts coated with or without Matrigel, respectively (8 μm pores, Corning).

Techniques: Co-Culture Assay, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Western Blot, Migration

Journal: Cancers

Article Title: Visfatin Mediates Malignant Behaviors through Adipose-Derived Stem Cells Intermediary in Breast Cancer

doi: 10.3390/cancers12010029

Figure Lengend Snippet: Visfatin-primed ADSCs promoted the tube formation of HUVEC. ( A ) HUVEC cells were co-cultured with visfatin-treated ADSCs or untreated ADSCs, noted as V50 and V100 or Ctrl, respectively, for three days. The HUVEC cells were collected from the co-culture and seeded in a matrix gel-coated 96-well plate. The tube formation of HUVEC was observed using a microscope. The length of branches was determined by using the ImageJ software. ( B ) After the three-day co-culture in the presence or absence of GDF15 neutralizing antibody (GDF15 Nab, 5 μg/mL), the HUVEC cells were collected from the co-culture for performing the tube formation assay. The experiments were performed in triplicate.

Article Snippet: For performing cell invasion or migration assay, the MDA-MB-231 cells collected from the co-culture system or treated with human recombinant GDF15 (10936-H01H, Sino Biological, Wayne, PA, USA) were suspended in serum-free DMEM and transferred to the 24-well transwell inserts coated with or without Matrigel, respectively (8 μm pores, Corning).

Techniques: Cell Culture, Co-Culture Assay, Microscopy, Software, Tube Formation Assay

Journal: Cancers

Article Title: Visfatin Mediates Malignant Behaviors through Adipose-Derived Stem Cells Intermediary in Breast Cancer

doi: 10.3390/cancers12010029

Figure Lengend Snippet: Visfatin-pretreated ADSCs (vADSCs) enhanced the tumor growth and metastasis in human breast cancer xenograft mouse model. ( A ) The nude mice were injected with mixture of MDA-MB-231 and untreated ADSCs (uADSCs) or vADSCs, noted as Ctrl or V50, respectively, to the mammary fat pads. The tumor volumes were measured every week after injection (Ctrl, n = 5; V50, n = 6). ( B ) After sacrificing the mice, the weight of the resected tumor was measured. ( C ) The expressions of GDF15, β-catenin, CD31, and pAKT in the tumor sections were detected by immunohistochemistry. The IHC score was calculated by multiplying the percentage of positive cells by the intensity and present as histogram. ( D ) The luciferase-expressing MDA-MB-231 were collected and injected into the tail vein of NOD/SCID mice after co-culturing with uADSCs or vADSCs, noted as Ctrl or V50, respectively (Ctrl, n = 8; V50, n = 8). The IVIS radiance signals of the mice were assessed at week 4. The representative images of high and low signal were shown. The statistical differences were calculated by t-test, *, p -value < 0.05; **, p -value < 0.01.

Article Snippet: For performing cell invasion or migration assay, the MDA-MB-231 cells collected from the co-culture system or treated with human recombinant GDF15 (10936-H01H, Sino Biological, Wayne, PA, USA) were suspended in serum-free DMEM and transferred to the 24-well transwell inserts coated with or without Matrigel, respectively (8 μm pores, Corning).

Techniques: Injection, Immunohistochemistry, Luciferase, Expressing

Journal: Cancers

Article Title: Visfatin Mediates Malignant Behaviors through Adipose-Derived Stem Cells Intermediary in Breast Cancer

doi: 10.3390/cancers12010029

Figure Lengend Snippet: The expressions of visfatin, GDF15, and pAKT in the specimens from breast cancer patients. ( A ) The expressions of visfatin, GDF15, and pAKT in breast cancer tissue microarray (n = 96) were detected by immunohistochemistry. The representative images of high expression levels (No. 1) and low expression levels (No. 2) were shown. The IHC score was calculated by multiplying the percentage of positive cells by the intensity, which was identified using HistoQuest Analysis Software. ( B ) The correlations between visfatin, GDF15, and pAKT according to the IHC score were calculated by using the online Pearson correlation coefficient calculator. ( C ) The correlation of serum levels of GDF15 and visfatin of breast cancer patients (n = 120) determined by ELISA was also calculated by using the online Pearson correlation coefficient calculator.

Article Snippet: For performing cell invasion or migration assay, the MDA-MB-231 cells collected from the co-culture system or treated with human recombinant GDF15 (10936-H01H, Sino Biological, Wayne, PA, USA) were suspended in serum-free DMEM and transferred to the 24-well transwell inserts coated with or without Matrigel, respectively (8 μm pores, Corning).

Techniques: Microarray, Immunohistochemistry, Expressing, Software, Enzyme-linked Immunosorbent Assay

Journal: Cancers

Article Title: Visfatin Mediates Malignant Behaviors through Adipose-Derived Stem Cells Intermediary in Breast Cancer

doi: 10.3390/cancers12010029

Figure Lengend Snippet: Visfatin mediates its effects both directly via cAbl/STAT3 and indirectly mediated by ADSCs via GDF15/AKT on promoting malignant behavior in breast cancer. Previously, we discovered visfatin mainly produced by adipocytes promoted breast cancer cells directly through activation of c-Abl and STAT3, which was blocked by Imatinib and Stattic inhibitor, respectively (black arrow). In this study, we showed that visfatin can act via an indirect pathway by priming ADSCs, which may be recruited from the adipose tissue to tumor site or generated from autologous fat transfer, to produce GDF15 that stimulated AKT activation in breast cancer cells to promote malignant behaviors (white arrow). The effect can be blocked by the treatment of GDF15 neutralizing Ab or Wortmannin inhibitor.

Article Snippet: For performing cell invasion or migration assay, the MDA-MB-231 cells collected from the co-culture system or treated with human recombinant GDF15 (10936-H01H, Sino Biological, Wayne, PA, USA) were suspended in serum-free DMEM and transferred to the 24-well transwell inserts coated with or without Matrigel, respectively (8 μm pores, Corning).

Techniques: Produced, Activation Assay, Generated

Journal: Redox Biology

Article Title: GDF15 nanotherapy ameliorates NLRP3-associated redox imbalance and cardiac injury in sepsis

doi: 10.1016/j.redox.2025.103897

Figure Lengend Snippet: Multifaceted validation of GDF15 changes in serum from SICM patients and their clinical associations. (A) GDF15 levels were quantified using the Luminex platform. (B) A volcano plot illustrated the gene expression distribution of GDF15 among differentially expressed genes (DEGs) in whole blood. (C) A heatmap displayed the expression profiles of GDF15 and inflammatory cytokines. (D) Serum GDF15 levels in patients. (E) Pearson correlation analysis demonstrated the association between GDF15 and SOFA score, as well as EF. (F) ROC curves were plotted to assess the diagnostic accuracy of GDF15 and SOFA score in identifying SICM. (G) Multivariate logistic regression analysis was performed to identify independent risk factors for the development of SICM in septic patients. ∗p < 0.05 indicates significant differences; ns: no significant differences.

Article Snippet: Enzyme-linked immunosorbent assays (ELISA) were conducted using commercial kits to quantify serum levels of human GDF15 (Proteintech, Cat# KE00108), mouse GDF15 (Proteintech, Cat# KE10082), and mouse IL-6 (MULTI SCIENCES, Cat# EK206).

Techniques: Biomarker Discovery, Luminex, Gene Expression, Expressing, Diagnostic Assay

Journal: Redox Biology

Article Title: GDF15 nanotherapy ameliorates NLRP3-associated redox imbalance and cardiac injury in sepsis

doi: 10.1016/j.redox.2025.103897

Figure Lengend Snippet: Upregulation of GDF15 in the SICM model. (A) A schematic workflow for the establishment of the SICM model in C57BL/6J mice via intraperitoneal injection of LPS or saline. (B) Cardiac contractile function parameters, including EF and FS. (C) Serum levels of GDF15 and IL-6. (D) Histopathological analysis of heart tissue, H&E staining (left) and immunohistochemical staining for Ly6G and CD68 (right). Black arrows indicate inflammatory cell infiltration; scale bar: 50 μm. (E) Western blot analysis of GDF15 protein expression in heart tissue. n = 4. (F) qPCR analysis of Gdf15 , Bnp , Il-1β , Il-6 , Icam-1 and Vcam- 1 mRNA levels in heart tissue. (G) Identification of GDF15-positive cells in single-cell RNA-sequencing dataset ( GSE190856 ). (H) qPCR analysis of Gdf15 and Il-1β , Il-6, Nos2, Ptgs2 mRNA expression in BMDM after LPS stimulation. ∗p < 0.05 indicates significant differences; n = 6 per group.

Article Snippet: Enzyme-linked immunosorbent assays (ELISA) were conducted using commercial kits to quantify serum levels of human GDF15 (Proteintech, Cat# KE00108), mouse GDF15 (Proteintech, Cat# KE10082), and mouse IL-6 (MULTI SCIENCES, Cat# EK206).

Techniques: Injection, Saline, Staining, Immunohistochemical staining, Western Blot, Expressing, RNA Sequencing

Journal: Redox Biology

Article Title: GDF15 nanotherapy ameliorates NLRP3-associated redox imbalance and cardiac injury in sepsis

doi: 10.1016/j.redox.2025.103897

Figure Lengend Snippet: GDF15 deficiency exacerbates LPS-induced SICM in mice. (A) Schematic workflow for the establishment of the SICM model in Gdf15 −/− mice. Gdf15 −/− mice were intraperitoneally injected with LPS or saline to induce SICM, with tissue samples collected 24 h post-injection for further analysis. (B) Echocardiographic assessment of EF and FS. (C) H&E staining of heart tissue, black arrows indicate inflammatory cell infiltration. scale bar: 50 μm. (D) CD68 immunofluorescence staining of heart tissue. Blue staining highlights nuclei, red staining identifies CD68 + macrophages; scale bar: 20 μm. (E) qPCR analysis of mRNA expression levels of Bnp , Il-1β, Il-6, and Mcp-1 in heart tissue. n = 6 per group.

Article Snippet: Enzyme-linked immunosorbent assays (ELISA) were conducted using commercial kits to quantify serum levels of human GDF15 (Proteintech, Cat# KE00108), mouse GDF15 (Proteintech, Cat# KE10082), and mouse IL-6 (MULTI SCIENCES, Cat# EK206).

Techniques: Injection, Saline, Staining, Immunofluorescence, Expressing

Journal: Redox Biology

Article Title: GDF15 nanotherapy ameliorates NLRP3-associated redox imbalance and cardiac injury in sepsis

doi: 10.1016/j.redox.2025.103897

Figure Lengend Snippet: MGP exerts anti-inflammatory effects via the MYPT1/AKT/YBX-1 signaling pathway. (A) IP-MS of BMDM to identify the interaction with GDF15. MYPT1 is marked in red. (B) Z-DOCK predicted the interaction domain between GDF15 and MYPT1. Pink represents GDF15, green represents MYPT1, and the boxed region indicates the binding domain. (C) Co-IP combined with Western blot analysis of GDF-15 and MYPT1 binding in macrophages after LPS treatment. (n = 3). (D) Immunofluorescence detection of co-localization between GDF15 (green) and MYPT1 (red), with blue staining for nuclei. Scale bar: 20 μm. (E) Protein expression levels of p -YBX-1, YBX-1, and p -AKT, AKT in BMDM after LPS and/or MGP treatment, with gray-scale intensity analysis of relative expression differences. (F) Representative immunofluorescence images of YBX-1 staining in BMDM after LPS and/or MGP treatment. Blue staining highlights nuclei, and red staining identifies YBX-1. Scale bar: 20 μm ∗p < 0.05, significantly different from control group. #p < 0.05, significantly different from LPS group. n = 6 per group.

Article Snippet: Enzyme-linked immunosorbent assays (ELISA) were conducted using commercial kits to quantify serum levels of human GDF15 (Proteintech, Cat# KE00108), mouse GDF15 (Proteintech, Cat# KE10082), and mouse IL-6 (MULTI SCIENCES, Cat# EK206).

Techniques: Protein-Protein interactions, Binding Assay, Co-Immunoprecipitation Assay, Western Blot, Immunofluorescence, Staining, Expressing, Control

Journal: Redox Biology

Article Title: GDF15 nanotherapy ameliorates NLRP3-associated redox imbalance and cardiac injury in sepsis

doi: 10.1016/j.redox.2025.103897

Figure Lengend Snippet: YBX-1 mediates GDF15-mediated transcriptional regulation of the NLRP3 pathway. (A) qPCR analysis of mRNA expression levels of Nlrp3, Asc , and Il-1β in LPS-stimulated BMDM after Si- Ybx-1 . (B) Western blot analysis of protein expression levels of NLRP3 and IL-1β in LPS-stimulated BMDM after YBX-1 knockdown. (C) qPCR analysis of mRNA expression levels of Nlrp3 and Il-1β in LPS and MGP-treated BMDM after YBX-1 knockdown. (D) Schematic diagram of the luciferase reporter plasmid for the Nlrp3 promoter. (E) Luciferase activity of pcDNA3.1-YBX-1 or empty vector-transfected cells. (F) Luciferase activity after LPS and MGP treatment. n = 6 per group.

Article Snippet: Enzyme-linked immunosorbent assays (ELISA) were conducted using commercial kits to quantify serum levels of human GDF15 (Proteintech, Cat# KE00108), mouse GDF15 (Proteintech, Cat# KE10082), and mouse IL-6 (MULTI SCIENCES, Cat# EK206).

Techniques: Expressing, Western Blot, Knockdown, Luciferase, Plasmid Preparation, Activity Assay, Transfection

Journal: Redox Biology

Article Title: GDF15 nanotherapy ameliorates NLRP3-associated redox imbalance and cardiac injury in sepsis

doi: 10.1016/j.redox.2025.103897

Figure Lengend Snippet: Mechanism of action of macrophage-biomimetic nanocarriers delivering GDF15 to target the YBX-1-NLRP3 axis in SICM. Macrophage-biomimetic nanocarriers loaded with rhGDF15 are targeted to inflammatory sites in the heart, enhancing local drug accumulation, while GDF15 binds to MYPT1 to inhibit YBX-1 phosphorylation and block its nuclear translocation, leading to reduced nuclear YBX-1 expression and decreased transcriptional activity of the Nlrp3 promoter, which suppresses NLRP3 inflammasome assembly and pro-inflammatory cytokine release such as IL-1β, ultimately alleviating macrophage inflammatory responses, myocardial cell injury, and improving cardiac function in SICM.

Article Snippet: Enzyme-linked immunosorbent assays (ELISA) were conducted using commercial kits to quantify serum levels of human GDF15 (Proteintech, Cat# KE00108), mouse GDF15 (Proteintech, Cat# KE10082), and mouse IL-6 (MULTI SCIENCES, Cat# EK206).

Techniques: Phospho-proteomics, Blocking Assay, Translocation Assay, Expressing, Activity Assay

Journal: Frontiers in Cardiovascular Medicine

Article Title: Serum GDF15, a Promising Biomarker in Obese Patients Undergoing Heart Surgery

doi: 10.3389/fcvm.2020.00103

Figure Lengend Snippet: Biomarker findings.

Article Snippet: In brief, paraffin sections were hydrated, antigens retrieved using Citrate buffer, blocked with 15% goat serum and incubated with anti- GDF15 primary antibody at 1:100 dilution (Santacruz Biotechnology) ( ).

Techniques: Biomarker Discovery

Journal: Frontiers in Cardiovascular Medicine

Article Title: Serum GDF15, a Promising Biomarker in Obese Patients Undergoing Heart Surgery

doi: 10.3389/fcvm.2020.00103

Figure Lengend Snippet: GDF15 significantly correlates with obesity but not with metabolic status of patients. (A) Levels of GDF15 in the plasma are increased in obese patients compared to non-obese patients. (B) GDF15 increased in obesity classes II and III, with a significant difference between normal and class III patients. (C) Circulating GDF15 also significantly correlates with body mass index (BMI), but not with glycated hemoglobin (HbA1c), triglycerides or age (D–F) . Data were analyzed using unpaired t -test, one-way ANOVA, Pearson's correlation and regression.

Article Snippet: In brief, paraffin sections were hydrated, antigens retrieved using Citrate buffer, blocked with 15% goat serum and incubated with anti- GDF15 primary antibody at 1:100 dilution (Santacruz Biotechnology) ( ).

Techniques: Clinical Proteomics

Journal: Frontiers in Cardiovascular Medicine

Article Title: Serum GDF15, a Promising Biomarker in Obese Patients Undergoing Heart Surgery

doi: 10.3389/fcvm.2020.00103

Figure Lengend Snippet: GDF15 protein expression in atrial tissues (AA) correspond with circulating GDF15. (A) Low to moderate cytoplasmic expression of GDF15 was observed in atrial tissues. (i) , Negative control. (ii) , GDF15 immunohistochemistry. (B) GDF15 showed weak protein expression in atrial tissue. GDF15 protein was normalized to whole protein stain. (C) Plasma GDF15 correlates with protein expression (i) , and with NT-proBNP, a marker for heart failure (ii) . Data were analyzed using Pearson's correlation and linear regression. For immunochemistry, outer magnification: 40X, scale bar: 50 μm. Arrows represent protein stain.

Article Snippet: In brief, paraffin sections were hydrated, antigens retrieved using Citrate buffer, blocked with 15% goat serum and incubated with anti- GDF15 primary antibody at 1:100 dilution (Santacruz Biotechnology) ( ).

Techniques: Expressing, Negative Control, Immunohistochemistry, Staining, Clinical Proteomics, Marker

Journal: Frontiers in Cardiovascular Medicine

Article Title: Serum GDF15, a Promising Biomarker in Obese Patients Undergoing Heart Surgery

doi: 10.3389/fcvm.2020.00103

Figure Lengend Snippet: Patients with GDF15 ≥75th percentile had a significantly worse prognosis. Using Kaplan Meier curves, followed by Log-Rank test, there was no significant difference in 2 year post-surgery survival between patients who were (i) non-obese/obese (ii) non-diabetic/diabetic (iv) or TG <75th percentile/≥75th percentile. However, patients with plasma GDF15 ≥75th percentile had a significantly poor post-surgery outcome compared to those with GDF15 <75th percentile (iii) .

Article Snippet: In brief, paraffin sections were hydrated, antigens retrieved using Citrate buffer, blocked with 15% goat serum and incubated with anti- GDF15 primary antibody at 1:100 dilution (Santacruz Biotechnology) ( ).

Techniques: Clinical Proteomics

Journal: International Journal of Molecular Sciences

Article Title: GDF15 Supports the Inflammatory Response of PdL Fibroblasts Stimulated by P. gingivalis LPS and Concurrent Compression

doi: 10.3390/ijms222413608

Figure Lengend Snippet: GDF15 promote the inflammatory response of HPdLF to lipopolysaccharides of P. gingivalis . ( a ) Quantitative analysis of GDF15 expression in HPdLF stimulated with 10 µg/ml P. gingivalis LPS for 24 hours. ( b ) Validation of GDF15 knockdown by quantitative analysis of GDF15 expression level in HPdLF treated with GDF15 siRNA ( GDF15 siR) or Control siRNA (Ctrl siR). ( c , d ) Analysis of adherent THP1 monocytic cells (green, Alexa488-gelabled) on HPdLF (blue, DAPI) after GDF15 knockdown as well as stimulation with P. gingivalis LPS ( c ). The relative number of THP1 cells is displayed per 100 HPdLF in ( d ). ( e – h ) Quantitative expression analysis of the inflammatory genes IL6 ( e ), IL8 ( f ), COX2 ( g ) and TNF α ( h ) in GDF15 -deficient HPdLF additionally stimulated with P. gingivalis LPS. ( i – l ) Analysis of secreted cytokines IL6 ( i ), IL8 ( j ), PGE2 ( k ) and TNFα ( l ) in GDF15 -deficient HPdLF additionally stimulated with P. gingivalis LPS. * p < 0.05; ** p < 0.01; *** p < 0.001 in relation to Ctrl ( a ) and Ctrl siR ( b – l ), ## p < 0.01; ### p < 0.001 in relation to GDF15 siR, § p < 0.05; §§§ p < 0.001 in relation to Ctrl siR + LPS; One-Way ANOVA and post hoc test (Tukey). Scale bars: 50 μm in ( c ). RNE, relative normalized expression.

Article Snippet: Human GDF15 siRNA oligos (50 nM, Santa Cruz Biotechnology, Dallas, TX, USA) or 50 nM BLOCK-iT Alexa Fluor red control siRNA (Thermo Fisher Scientific, Carlsbad, CA, USA) were applied for five hours in OptiMem I-reduced serum medium (Thermo Fisher Scientific, Carlsbad, CA, USA) containing 100 U/mL penicillin and 100 μg/mL streptomycin.

Techniques: Expressing, Biomarker Discovery, Knockdown, Control

Journal: International Journal of Molecular Sciences

Article Title: GDF15 Supports the Inflammatory Response of PdL Fibroblasts Stimulated by P. gingivalis LPS and Concurrent Compression

doi: 10.3390/ijms222413608

Figure Lengend Snippet: The pro-inflammatory response of HPdLF to mechanical and bacterial stimuli is partially modulated by GDF15. ( a – d ) Quantitative expression analysis of the inflammatory genes IL6 ( a ), IL8 ( b ), COX2 ( c ) and TNFα ( d ) in HPdLF treated with GDF15 siRNA ( GDF15 siR) or Control siRNA (Ctrl siR), stimulated for six h with compressive force (+CF, white bars) and lipopolysaccharides of P. gingivalis (+LPS, black bars). The expression levels are displayed as fold change (FC) to Ctrl siR-treated cells. ( e – h ) Analysis of secreted cytokines IL6 ( e ), IL8 ( f ), PGE2 ( g ), and TNFα ( h ) in HPdLF under previous conditions. ( i , j ) Analysis of adherent THP1 monocytic cells (green, Alexa488-gelabled) on mechanically stressed GDF15 -deficient HPdLF (blue, DAPI) in relation to the control under previous conditions ( i ). The relative number of THP1 cells is displayed per 100 HPdLF in ( j ). * p < 0.05; ** p < 0.01; *** p < 0.001 in relation to Ctrl siR +CF; # p < 0.05; ## p < 0.01; ### p < 0.001 in relation to GDF15 siR +CF, § p < 0.05; §§ p < 0.01; §§§ p < 0.001 in relation to Ctrl siR +CF +LPS. One-Way ANOVA and post hoc test (Tukey). Scale bars: 50 μm in ( c ).

Article Snippet: Human GDF15 siRNA oligos (50 nM, Santa Cruz Biotechnology, Dallas, TX, USA) or 50 nM BLOCK-iT Alexa Fluor red control siRNA (Thermo Fisher Scientific, Carlsbad, CA, USA) were applied for five hours in OptiMem I-reduced serum medium (Thermo Fisher Scientific, Carlsbad, CA, USA) containing 100 U/mL penicillin and 100 μg/mL streptomycin.

Techniques: Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: GDF15 Supports the Inflammatory Response of PdL Fibroblasts Stimulated by P. gingivalis LPS and Concurrent Compression

doi: 10.3390/ijms222413608

Figure Lengend Snippet: qPCR primer sequences of human genes indicated in 5′-3′ direction. bp, base pairs. Length, amplicon length.

Article Snippet: Human GDF15 siRNA oligos (50 nM, Santa Cruz Biotechnology, Dallas, TX, USA) or 50 nM BLOCK-iT Alexa Fluor red control siRNA (Thermo Fisher Scientific, Carlsbad, CA, USA) were applied for five hours in OptiMem I-reduced serum medium (Thermo Fisher Scientific, Carlsbad, CA, USA) containing 100 U/mL penicillin and 100 μg/mL streptomycin.

Techniques: Amplification, Sequencing, Binding Assay

Journal: Scientific Reports

Article Title: Growth differentiation factor-15: a p53- and demethylation-upregulating gene represses cell proliferation, invasion, and tumorigenesis in bladder carcinoma cells

doi: 10.1038/srep12870

Figure Lengend Snippet: Expressions of GDF15 in HT1376 cells stably transfected with pcDNA3 (HT-DNA) or pcDNA-GDF15 (HT-GDF15) expression vector were determined by immunoblotting assays ( a ) and ELISA ( b ). Data are expressed as mean-fold (±S.E.; n = 3) in relation to the HT-DNA cell group and the mean (±S.E.; n = 6) of the GDF15 levels. Proliferations of HT-DNA (white circle) and HT-GDF15 (black circle) cells were determined according to the incorporation of 3 H-thymidine ( c ) and MTS assays ( d ). Each point on the curve represents the mean-percentage (±S.E.; n = 6) of that on day 1. ( e ) The invasive ability of cells was determin e d by in vitro matrigel invasion assays. Data are presented as mean-percentage (±S.E.; n = 3) in relation to that of the HT-DNA cell group. ( f ) Nude mice were inoculated subcutaneously with HT-DNA (black circle) or HT-GDF15 (white circle) cells. At the indicated days, tumor size was measured using vernier calipers, and results are presented as tumor size in mm 3 (±S.E.). ( g ) Blood samples were collected from experimental animals by cardiocentesis immediately after sacrificed, and GDF15 levels were determined by ELISA. Data is presented as mean (±S.E.; n = 6) of the GDF15 levels. (*P < 0.05, + P < 0.01).

Article Snippet: We purchased fetal calf serum (FCS) from HyClone (Logan, UT, USA), RPMI 1640 media from Life Technologies (Rockville, MD, USA), recombinant human GDF15 (rhGDF15) from PeproTech (Rocky Hill, NJ, USA), matrigel from BD Biosciences (Bedford, MA, USA), and doxorubicin, camptothecin, and 5′-aza-2′deoxycytidine from Sigma (St. Louis, MO, USA).

Techniques: Stable Transfection, Transfection, Expressing, Plasmid Preparation, Western Blot, Enzyme-linked Immunosorbent Assay, In Vitro

Journal: Scientific Reports

Article Title: Growth differentiation factor-15: a p53- and demethylation-upregulating gene represses cell proliferation, invasion, and tumorigenesis in bladder carcinoma cells

doi: 10.1038/srep12870

Figure Lengend Snippet: Expressions of GDF15 in mock-knockdown HT1376 (HT-COLsi) and GDF15 knockdown HT1376 (HT-GDF15si) cells were determined by immunoblotting ( a , top) and RT-qPCR ( a , bottom) assays. Data are expressed as mean-fold of the GDF15 levels (±S.E.; n = 3) in relation to the HT-COLsi cell group. Proliferations of HT-GDF15si (white circle) and HT-COLsi (black circle) cells were determined according to the incorporation of 3 H-thymidine ( b ) and MTS assays ( c ). Each point on the curve represents the mean-percentage (±S.E.; n = 6) of that on day 1. ( d ) Invasive ability of cells was determined by the in vitro matrigel invasion assays. Data are presented as mean-percentage (±S.E.) in relation to the HT-COLsi cell group. ( e ) Nude mice were inoculated subcutaneously with HT-COLsi (black circle) or HT-GDF15si (white circle) cells. Tumor size as measured using vernier calipers. Results are presented as tumor size in mm 3 (±S.E.), which measured at the indicated time intervals. (*P < 0.05, + P < 0.01).

Article Snippet: We purchased fetal calf serum (FCS) from HyClone (Logan, UT, USA), RPMI 1640 media from Life Technologies (Rockville, MD, USA), recombinant human GDF15 (rhGDF15) from PeproTech (Rocky Hill, NJ, USA), matrigel from BD Biosciences (Bedford, MA, USA), and doxorubicin, camptothecin, and 5′-aza-2′deoxycytidine from Sigma (St. Louis, MO, USA).

Techniques: Western Blot, Quantitative RT-PCR, In Vitro

Journal: Scientific Reports

Article Title: Growth differentiation factor-15: a p53- and demethylation-upregulating gene represses cell proliferation, invasion, and tumorigenesis in bladder carcinoma cells

doi: 10.1038/srep12870

Figure Lengend Snippet: Expressions of GDF15 in T24 cells transfected with pcDNA3 (T24-DNA) or pcDNA-GDF15 (T24-GDF15) expression vector were determined by immunoblotting assays ( a , top), RT-qPCR assays ( a , bottom) and ELISA ( b ). Data are expressed as mean-fold (±S.E.; n = 3) in relation to the T24-DNA cell group and the mean (±S.E.; n = 6) of the GDF15 levels. Cell proliferations in T24-DNA (white circle) and T24-GDF15 (black circle) were determined according to 3 H-thymidine incorporation ( c ) and MTS assays ( d ). Each point on the curve represents the mean-percentage (±S.E.; n = 6) of that on day 1. ( e ) The invasive ability of cells was determined by the in vitro matrigel invasion assays. Data are presented as the mean-percentage (±S.E.) in relation to that of the T24-DNA cell group. (*P < 0.05, + P < 0.01).

Article Snippet: We purchased fetal calf serum (FCS) from HyClone (Logan, UT, USA), RPMI 1640 media from Life Technologies (Rockville, MD, USA), recombinant human GDF15 (rhGDF15) from PeproTech (Rocky Hill, NJ, USA), matrigel from BD Biosciences (Bedford, MA, USA), and doxorubicin, camptothecin, and 5′-aza-2′deoxycytidine from Sigma (St. Louis, MO, USA).

Techniques: Transfection, Expressing, Plasmid Preparation, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, In Vitro

Journal: Science signaling

Article Title: Inflammation induces stress erythropoiesis through heme-dependent activation of SPI-C

doi: 10.1126/scisignal.aap7336

Figure Lengend Snippet: (A) Schematic of the LPS-zymosan model of sterile inflammation. (B) Splenocytes were harvested at the indicated time points and plated at a concentration of 1 × 105 cells per well in the presence of GDF15, BMP4, SCF, SHH, and EPO at low O2 to induce the formation of stress BFU-Es. Stress BFU-Es were scored after 5 days by staining with benzidine. Data represent the mean ± SEM, one-way ANOVA with Dunnett’s multiple comparison test, n = 4 to 11 mice per time point. (C) Flow cytometry analysis of splenocytes was performed at 0 and 36 hours after treatment with zymosan. Cells were gated on Kit+ cells, and frequencies of CD34+ and CD133+ populations are shown in representative images. n = 3 to 5 mice per time point. (D and E) RNA was isolated from splenocytes at the indicated time points after zymosan injection, and relative expression of Gdf15 and Bmp4 compared to 18S was determined by quantitative PCR. Data represent the mean ± SEM, one-way ANOVA with Dunnett’s multiple comparison test, n = 3 to 10 mice per time point. (F) Splenocytes harvested from mice at the indicated time points after zymosan treatment were plated under conditions to induce stress BFU-Es and stained with benzidine and counted after 5 days. Data represent the mean ± SEM, one-way ANOVA with Dunnett’s multiple comparison test, n = 3 to 9 mice per time point. (G) Protein was isolated from kidneys at the indicated time points after zymosan treatment and Western blotted for HIF-2α and β-actin. HIF-2α abundance relative to β-actin was calculated using ImageJ. Data represent the mean ± SEM, one-way ANOVA with Dunnett’s multiple comparison test, n = 3 mice per time point. (H) Quantification of Epo expression relative to 18S in RNA isolated from kidneys and abundance of EPO in serum at the indicated time points after zymosan treatment. Data represent the mean ± SEM, one-way ANOVA with Dunnett’s multiple comparison test, n = 2 to 5 mice per time point (mRNA) and 2 to 8 mice per time point (protein). *P < 0.05, **P < 0.01, and ***P < 0.005.

Article Snippet: Briefly, SEEM is composed of Iscove’s modified Dulbecco’s media containing 10% fetal bovine serum, insulin (10 μg/ml), holo-transferrin (200 μg/ml), 2 mM L-glutamine, ciprofloxacin (10 μg/ml), 1% bovine serum albumin (BSA), 2-mercaptoethanol (7 μl/liter), GDF15 (30 ng/ml, Biomatik), BMP4 (15 ng/ml, R&D Systems), sonic hedgehog (SHH) (25 ng/ml, GoldBio), and stem cell factor (SCF) (50 ng/ml, Gold-Bio).

Techniques: Concentration Assay, Staining, Flow Cytometry, Isolation, Injection, Expressing, Real-time Polymerase Chain Reaction, Western Blot

Journal: Bioengineered

Article Title: Protective effect of growth differentiation factor 15 in sepsis by regulating macrophage polarization and its mechanism

doi: 10.1080/21655979.2022.2059903

Figure Lengend Snippet: Expressions of macrophage markers and inflammatory factor mRNA

Article Snippet: Dulbecco’s modified eagle medium (DMEM), a high-glucose medium containing 10% fetal bovine serum (FBS; HyClone, USA); collagenase IV (2 mg/ml; Worthington, Lakewood, NJ, USA); neutral protease II (1.2 U/mL; Sigma-Aldrich, USA); hyaluronidase (Sigma-Aldrich, USA); mouse lymphocyte isolate, Ficoll (Sigma-Aldrich, USA); allophycocyanin anti-F4/80 (BioLegend, USA); anti-CD11b antibody (ab184308; Abcam, UK); Alexa Fluor® 488 anti–CD206 (BioLegend, USA); fluorescein isothiocyanate anti-CD80 (Becton, Dickinson and Company, USA); Ly6C antibody (ab24973; Abcam, UK); Ly6G antibody (ab25377; Abcam, UK); MHC-II antibody (ab180779; Abcam, UK); PI3K inhibitor, LY294002, and corresponding phosphorylated antibody (Cell Signaling Technology, Inc., USA); and recombinant GDF15 protein (rGDF15; PeproTech, USA) were used in this study.

Techniques:

Journal: Bioengineered

Article Title: Protective effect of growth differentiation factor 15 in sepsis by regulating macrophage polarization and its mechanism

doi: 10.1080/21655979.2022.2059903

Figure Lengend Snippet: Protein expression of p-PI3K and p-Akt in the PI3K/AKT signaling pathway of macrophages of each group

Article Snippet: Dulbecco’s modified eagle medium (DMEM), a high-glucose medium containing 10% fetal bovine serum (FBS; HyClone, USA); collagenase IV (2 mg/ml; Worthington, Lakewood, NJ, USA); neutral protease II (1.2 U/mL; Sigma-Aldrich, USA); hyaluronidase (Sigma-Aldrich, USA); mouse lymphocyte isolate, Ficoll (Sigma-Aldrich, USA); allophycocyanin anti-F4/80 (BioLegend, USA); anti-CD11b antibody (ab184308; Abcam, UK); Alexa Fluor® 488 anti–CD206 (BioLegend, USA); fluorescein isothiocyanate anti-CD80 (Becton, Dickinson and Company, USA); Ly6C antibody (ab24973; Abcam, UK); Ly6G antibody (ab25377; Abcam, UK); MHC-II antibody (ab180779; Abcam, UK); PI3K inhibitor, LY294002, and corresponding phosphorylated antibody (Cell Signaling Technology, Inc., USA); and recombinant GDF15 protein (rGDF15; PeproTech, USA) were used in this study.

Techniques: Expressing

Journal: Bioengineered

Article Title: Protective effect of growth differentiation factor 15 in sepsis by regulating macrophage polarization and its mechanism

doi: 10.1080/21655979.2022.2059903

Figure Lengend Snippet: Protein expression of p-PI3K and p-Akt in the PI3K/AKT signaling pathway of macrophages of each group (Figure 6a: * p < 0.05,LPS+GDF15 group vs. LPS group;*# p < 0.05,LPS+GDF15+ LY294002 groupvs.LPS+GDF15 group).

Article Snippet: Dulbecco’s modified eagle medium (DMEM), a high-glucose medium containing 10% fetal bovine serum (FBS; HyClone, USA); collagenase IV (2 mg/ml; Worthington, Lakewood, NJ, USA); neutral protease II (1.2 U/mL; Sigma-Aldrich, USA); hyaluronidase (Sigma-Aldrich, USA); mouse lymphocyte isolate, Ficoll (Sigma-Aldrich, USA); allophycocyanin anti-F4/80 (BioLegend, USA); anti-CD11b antibody (ab184308; Abcam, UK); Alexa Fluor® 488 anti–CD206 (BioLegend, USA); fluorescein isothiocyanate anti-CD80 (Becton, Dickinson and Company, USA); Ly6C antibody (ab24973; Abcam, UK); Ly6G antibody (ab25377; Abcam, UK); MHC-II antibody (ab180779; Abcam, UK); PI3K inhibitor, LY294002, and corresponding phosphorylated antibody (Cell Signaling Technology, Inc., USA); and recombinant GDF15 protein (rGDF15; PeproTech, USA) were used in this study.

Techniques: Expressing

Journal: Bioengineered

Article Title: Protective effect of growth differentiation factor 15 in sepsis by regulating macrophage polarization and its mechanism

doi: 10.1080/21655979.2022.2059903

Figure Lengend Snippet: Expression of macrophage phenotypic surface markers in mouse peritoneal cavity

Article Snippet: Dulbecco’s modified eagle medium (DMEM), a high-glucose medium containing 10% fetal bovine serum (FBS; HyClone, USA); collagenase IV (2 mg/ml; Worthington, Lakewood, NJ, USA); neutral protease II (1.2 U/mL; Sigma-Aldrich, USA); hyaluronidase (Sigma-Aldrich, USA); mouse lymphocyte isolate, Ficoll (Sigma-Aldrich, USA); allophycocyanin anti-F4/80 (BioLegend, USA); anti-CD11b antibody (ab184308; Abcam, UK); Alexa Fluor® 488 anti–CD206 (BioLegend, USA); fluorescein isothiocyanate anti-CD80 (Becton, Dickinson and Company, USA); Ly6C antibody (ab24973; Abcam, UK); Ly6G antibody (ab25377; Abcam, UK); MHC-II antibody (ab180779; Abcam, UK); PI3K inhibitor, LY294002, and corresponding phosphorylated antibody (Cell Signaling Technology, Inc., USA); and recombinant GDF15 protein (rGDF15; PeproTech, USA) were used in this study.

Techniques: Expressing