Journal: Pediatric research

Article Title: Circulating growth-and-differentiation factor-15 in early life: relation to prenatal and postnatal growth and adiposity measurements.

doi: 10.1038/s41390-019-0633-z

Figure Lengend Snippet: Fig. 1 GDF15 longitudinal data in the study population (70 AGA and 33 SGA). The upper and lower limits of the gray zone correspond to a Z-score of +1 and −1, respectively, in healthy adults (n = 18; age, 41.4 ± 0.7 years; BMI, 25.1 ± 0.5 kg/m2; circulating GDF15, 355 ± 18 pg/mL (mean ± SEM)). *P < 0.001 between values at birth and those at 4, 12 and 24 months; #P < 0.01 for AGA vs. SGA

Article Snippet: GDF15 levels were measured in duplicate using a specific human GDF15 ELISA kit (R&D Systems, Minneapolis), according to the manufacturer’s instructions; intra- and inter-assay CVs were <6%.

Techniques:

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: All bladder cells used in this study were serum starved for 24 hours subsequently incubated in RPMI media containing 10% FCS for another 24 hours. ( a ) Cell proteins were then lysed for immunoblotting assay. ( b ) Total RNA was extracted from cells for the RT-qPCR assay. Data are presented as mean-fold (±S.E.; n = 3) in relation to that of the RT4 cell group. ( c ) Conditioned media was collected for ELISA in order to determine the level of GDF15 secretion in the various bladder carcinoma cells. Data is presented as the mean (±S.E.; n = 6) of the GDF15 levels. ( d ) Proliferation rates in HT1376 (black circle) and T24 (white circle) cells treated with various concentrations of GDF15 were determined by 3 H-thymidine incorporation assays. Each point on the curve represents the mean-percentage (±S.E.; n = 6) relative to solvent-treated group (*P < 0.05, + P < 0.01).

Article Snippet: FAM dye-labeled TaqMan MGB probes as well as PCR primers for human GDF15 (Hs00171132_m1), NDRG1 (Hs00608387_m1), NDRG2 (Hs01045115_m1), NDRG3 (Hs00223890_m1), MASPIN (Hs00985283_m1), E-cadherin (Hs00170423_m1), N-cadherin (Hs00983062_m1), and β-actin (Hs01060665_g1) were purchased from Applied Biosystems.

Techniques: Incubation, Western Blot, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Solvent

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: ( a ) HT1376 (left) and T24 (right) cells were transiently overexpressed with p53 for 72 hours. The levels of GDF15 and p53 expressions were determined by immunoblotting assays. ( b ) The GDF15 report vector was co-transfected with various concentrations of p53 expression vector into HT1376 cells for 72 hours. Data are expressed as the mean percentage ± S.E. (n = 6) of luciferase activity relative to mock-transfected groups. Expressions of GDF15 and p53 in RT4 cells following doxorubicin ( c ) or camptothecin ( d ) treatments were determined by immunoblotting assays. ( e ) GDF15 secretion in RT4 cells following camptothecin treatments was determined by ELISA. Data are expressed as mean (±S.E.; n = 6) of the GDF15 levels. Total RNAs were extracted from doxorubicin treated ( f ) and camptothecin treated RT4 ( g ) cells for RT-qPCR assays. T24 cells were treated with various concentrations of 5-Aza-2′-deoxycytidine for 48 hours and then GDF15 expression was determined by immunoblotting ( h ), ELISA ( i ), and RT-qPCR assays ( j ). Data are expressed as the mean-fold ± S.E. (n = 3) relative to solvent-treated groups and mean (±S.E.; n = 6) of the GDF15 levels. (*P < 0.05, + P < 0.01).

Article Snippet: FAM dye-labeled TaqMan MGB probes as well as PCR primers for human GDF15 (Hs00171132_m1), NDRG1 (Hs00608387_m1), NDRG2 (Hs01045115_m1), NDRG3 (Hs00223890_m1), MASPIN (Hs00985283_m1), E-cadherin (Hs00170423_m1), N-cadherin (Hs00983062_m1), and β-actin (Hs01060665_g1) were purchased from Applied Biosystems.

Techniques: Western Blot, Plasmid Preparation, Transfection, Expressing, Luciferase, Activity Assay, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Solvent

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: FAM dye-labeled TaqMan MGB probes as well as PCR primers for human GDF15 (Hs00171132_m1), NDRG1 (Hs00608387_m1), NDRG2 (Hs01045115_m1), NDRG3 (Hs00223890_m1), MASPIN (Hs00985283_m1), E-cadherin (Hs00170423_m1), N-cadherin (Hs00983062_m1), and β-actin (Hs01060665_g1) were purchased from Applied Biosystems.

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: FAM dye-labeled TaqMan MGB probes as well as PCR primers for human GDF15 (Hs00171132_m1), NDRG1 (Hs00608387_m1), NDRG2 (Hs01045115_m1), NDRG3 (Hs00223890_m1), MASPIN (Hs00985283_m1), E-cadherin (Hs00170423_m1), N-cadherin (Hs00983062_m1), and β-actin (Hs01060665_g1) were purchased from Applied Biosystems.

Techniques: Knockdown, 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: FAM dye-labeled TaqMan MGB probes as well as PCR primers for human GDF15 (Hs00171132_m1), NDRG1 (Hs00608387_m1), NDRG2 (Hs01045115_m1), NDRG3 (Hs00223890_m1), MASPIN (Hs00985283_m1), E-cadherin (Hs00170423_m1), N-cadherin (Hs00983062_m1), and β-actin (Hs01060665_g1) were purchased from Applied Biosystems.

Techniques: Transfection, Expressing, Plasmid Preparation, Western Blot, Quantitative RT-PCR, 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: ( a ) Differences in expressions of NDRG1, NDRG2, NDRG3, and MASPIN between HT-DNA and HT-GDF15 cells were determined by immunoblotting assays. Data of quantitative analysis are expressed as the intensity of the protein bands produced from the target gene/β-actin (±S.E.; n = 3) of HT-GDF15 cells relative to that of the HT-DNA group. ( b ) The reporter vectors of NDRG1, NDRG3, and MASPIN were cotransfected with various concentrations of GDF15 expression vectors into HT1376 cells for 72 hours. Data are expressed as the mean percentage ± S.E. (n = 6) relative to the mock-transfected groups. Expressions of GDF15, NDRG1, NDRG2, NDRG3, and MASPIN in mock-knockdown HT1376 (HT-COLsi) and GDF15-knockdwon HT1376 (HT-GDF15si) cells were determined by immunoblotting ( c ) and RT-qPCR ( d ) assays. Data are presented as mean-fold (±S.E.) in relation to the HT-COLsi cell group. ( e ) Differences in the expressions of GDF15, NDRG1, NDRG2, NDRG3, and MASPIN genes between T24-DNA and T24-GDF15 cells were determined by RT-qPCR assays. Data are presented as mean-fold (±S.E.) in relation to the T24-DNA cell group. Differences in the expressions of NDRG1, NDRG2, NDRG3, and MASPIN genes following treatments with various concentrations of rhGDF15 were determined by immunoblotting ( f ) and RT-qPCR ( h ) assays. Data of quantitative analysis are expressed as the intensity of the protein bands produced from the target gene/β-actin (±S.E.; n = 3) relative to that of the solvent-control group ( g ). (*P < 0.05, + P < 0.01).

Article Snippet: FAM dye-labeled TaqMan MGB probes as well as PCR primers for human GDF15 (Hs00171132_m1), NDRG1 (Hs00608387_m1), NDRG2 (Hs01045115_m1), NDRG3 (Hs00223890_m1), MASPIN (Hs00985283_m1), E-cadherin (Hs00170423_m1), N-cadherin (Hs00983062_m1), and β-actin (Hs01060665_g1) were purchased from Applied Biosystems.

Techniques: Western Blot, Produced, Expressing, Transfection, Knockdown, Quantitative RT-PCR, Solvent, Control

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: ( a ) Expression levels of SNAIL, SLUG, E-cadherin, and N-cadherin in HT-GDF15si and HT-COLsi cells were determined by immunoblotting a ssays. ( b ) Expressions of GDF15, E-cadherin, and N-cadherin in HT-GDF15si (white bars) and HT-COLsi (black bars) cells were determined by RT-qPCR assays. Data are presented as mean-fold (±S.E.) in relation to the HT-COLsi cell group. ( c ) Distribution of F-actin (red) between HT-GDF15si and HT-COLsi cells was determined by immunofluorescence staining. DAPI (blue) was applied to stain the nucleus. (*P < 0.05, + P < 0.01).

Article Snippet: FAM dye-labeled TaqMan MGB probes as well as PCR primers for human GDF15 (Hs00171132_m1), NDRG1 (Hs00608387_m1), NDRG2 (Hs01045115_m1), NDRG3 (Hs00223890_m1), MASPIN (Hs00985283_m1), E-cadherin (Hs00170423_m1), N-cadherin (Hs00983062_m1), and β-actin (Hs01060665_g1) were purchased from Applied Biosystems.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Growth differentiation factor 15 (GDF15) expression is elevated in gastric cancer and high serum GDF15 level is a poor prognostic factor. (A) GDF15 gene expression level in gastric cancer patients illustrated with boxplots by the Gene Expression Profiling Interactive Analysis (GEPIA) online database. (B) The Kaplan–Meier plotter online database was used to analyze the clinical effect of GDF15 gene expression in gastric cancer patients ( http://kmplot.com/analysis/ ). (C) Clinical effect of GDF15 serum levels (≥upper quartile 1066.79 ng/mL vs.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Expressing, Gene Expression

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Clinical characteristics of patients with gastric cancer with different growth differentiation factor 15 (GDF15) expression of tumor tissues

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Expressing

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Clinical characteristics of patients with gastric cancer with low or high serum growth differentiation factor 15 (GDF15) levels

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques:

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Growth differentiation factor 15 (GDF15) expression is essential for cell proliferation and migration of gastric cancer cells. (A, C) siGDF15 (180 pmol for 3 × 10 5 cells in a 6‐cm dish for 48 h) or (B, D) pcDNA‐GDF15 (pGDF15, 6 μg for 3 × 10 5 cells in a 6‐cm dish for 12 h, followed by replacement of fresh medium for a total of 48 h) were used to knockdown or overexpress GDF15. Efficiencies of knockdown and overexpression were analyzed by quantitative real‐time PCR. (A, B) After transfection with siGDF15 or pGDF15, cells were reseeded with a density of 3000 cells per well in a 96‐well plate. Cell proliferation was analyzed with sulforhodamine B (SRB) assay. (C, D) After transfection with siGDF15 or pGDF15, cells were reseeded with a density of 1 × 10 5 cells per Transwell insert. Cell migration was determined by Transwell migration assay (siGDF15: AGS, NUGC‐3, and TSGH9201 for 12, 16, and 24 h migration, respectively; magnification, 200×) (pGDF15: AGS, NUGC‐3, and TSGH9201 for 8, 12, and 24 h migration, respectively; magnification, 100×). Graph is presented as mean ± SEM ( n ≥ 3). *Significant vs. individual control.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Expressing, Migration, Knockdown, Over Expression, Real-time Polymerase Chain Reaction, Transfection, Sulforhodamine B Assay, Transwell Migration Assay, Control

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Growth differentiation factor 15 (GDF15) contributes to cisplatin resistance in human gastric cancer cells. (A–C) GDF15 (A) gene and (B) protein expressions and (C) released GDF15 level between parental (P) and cisplatin‐resistant (CisR) gastric cancer cells were analyzed with quantitative real‐time PCR, western blotting, and ELISA assays, respectively. (D, E) Cisplatin sensitivity (48 h) of the gastric cancer cells was assessed using (D) sulforhodamine B (SRB) assay and (E) propidium iodide (PI) exclusion assay. (F–H) Effects of (F) GDF15 neutralizing Ab (GDF15 NAb), (G) recombinant human GDF15 (rhGDF15), and (H) GDF15 overexpression on sensitivity of cisplatin were evaluated with SRB assay. G, GDF15 plasmid; V, empty vector. Quantitative real‐time PCR and western blotting were used to validate the efficiencies of GDF15 knockdown or overexpression, respectively. Graph is presented by mean ± SEM ( n ≥ 3). *Significant vs. individual control. ** , ***Significant, rhGDF15 (20 and 50 ng/mL) vs. individual control.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Sulforhodamine B Assay, Exclusion Assay, Recombinant, Over Expression, Plasmid Preparation, Knockdown, Control

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Growth differentiation factor 15 (GDF15)‐upregulated xCT expression through the eukaryotic initiation factor 2α (eIF2α)‐activating transcription factor 4 (ATF4) pathway enhances intracellular glutathione (GSH) levels in cisplatin‐resistant gastric cancer cells. (A) Knockdown efficiency was validated using western blotting. (B) Effects of siGDF15 and glial cell‐derived neurotrophic factor family receptor a‐like siRNA (siGFRAL) on GSH levels were evaluated using the GSH detection kit. (C, D) After treatment of GDF15‐knockdown cisplatin‐resistant (CisR) cells with cisplatin (24 h), intracellular and mitochondrial reactive oxygen species were evaluated with (C) dichlorodihydro‐fluorescein (DCF) and (D) MitoSox Red using flow cytometry. (E) GDF15 and xCT gene expressions were evaluated using quantitative real‐time PCR. (F) Protein expression of GDF15 and the eIF2α‐xCT pathway were evaluated using western blotting. (G) After transfections with different xCT promoters (WT, antioxidant‐responsive element [ARE]‐mutant, and amino acid response element [AARE]‐mutant), the cells were further transfected with siGDF15. Graph is presented as mean ± SEM ( n ≥ 3). *Significant vs. individual control. **Significant vs. WT/ARE‐mutant‐xCT promoters.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Expressing, Knockdown, Western Blot, Derivative Assay, Flow Cytometry, Real-time Polymerase Chain Reaction, Transfection, Mutagenesis, Control

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Eukaryotic initiation factor 2α (eIF2α)‐activating transcription factor 4 (ATF4)‐xCT‐elevated glutathione (GSH) contributes to growth differentiation factor 15 (GDF15)‐mediated cisplatin resistance in gastric cancer cells. (A, B, D) Cells were transfected with pcDNA‐GDF15. G, GDF15 plasmid; V, empty vector. (A, D) Gene expressions of GDF15 and xCT were evaluated using quantitative real‐time PCR. (B) The eIF2α‐ATF4‐xCT pathway was evaluated using western blotting. (C) After transfection with different types of xCT promoters, HEK293T cells were further transfected with pcDNA‐GDF15. (D) After overexpression of GDF15, the effects of sulfasalazine (SSA, 350 μM) and buthionine sulfoximine (BSO, 0.5 mM) on cisplatin sensitivity (48 h) were evaluated with propidium iodide (PI) exclusion assay. Graph is presented as mean ± SEM ( n ≥ 3). *Significant vs. individual control. **Significant vs. WT/antioxidant‐responsive element (ARE)‐mutant xCT promoters. # Significant vs. pcDNA. ## Significant vs. cisplatin treatment. AARE, amino acid response element; Con, control.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction, Western Blot, Over Expression, Exclusion Assay, Control, Mutagenesis

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Growth differentiation factor 15 (GDF15)/ glial cell‐derived neurotrophic factor family receptor a‐like (GFRAL)‐mediated signaling in cisplatin‐resistant gastric cancer cells could be through general control nonderepressible 2 (GCN2). (A, B) siGDF15 and siGFRAL were transfected into (A) AGS cisplatin‐resistant (CisR) and (B) NUGC‐3CisR cells. (C) AGSCisR and (D) NUGC‐3CisR cells were treated with SPP86 (5 μΜ) for 24 h. Upstream regulators of the eukaryotic initiation factor 2α (eIF2α) and eIF2α‐activating transcription factor 4 (ATF4)‐xCT pathways were analyzed using western blotting. Graph is presented as mean ± SEM ( n ≥ 3). *Significant vs. individual control (Con). PERK, PKR‐like endoplasmic reticulum kinase; PKR, protein kinase R.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Derivative Assay, Control, Transfection, Western Blot

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: General control nonderepressible 2 (GCN2) is responsible for growth differentiation factor 15 (GDF15)‐mediated glial cell‐derived neurotrophic factor family receptor a‐like (GFRAL)‐eukaryotic initiation factor 2α (eIF2α)‐activating transcription factor 4 (ATF4)‐xCT signaling and cisplatin resistance. (A, B, D) After GDF15 overexpression by pcDNA‐GDF15 (G, pcDNA‐GDF15; V, pcDNA alone), cells were treated with siRNAs against (A) protein kinase R (PKR), (B) heme‐regulated eIF2α kinase (HRI), and (D) GCN2 for 48 h. The effect of siRNAs against PKR, HRI, and GCN2 on GDF15‐mediated eIF2α‐ATF4‐xCT regulation was assessed using western blotting. (C) Effects of siHRI and siPKR on cisplatin resistance in cisplatin‐resistant (CisR) cells was evaluated with sulforhodamine B (SRB) assay. Graph is presented as mean ± SEM ( n ≥ 3). *Significant vs. individual control. **Significant vs. siScr with GDF15 overexpression.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Control, Derivative Assay, Over Expression, Western Blot, Sulforhodamine B Assay

Journal: Cancer Science

Article Title: Growth differentiation factor 15 induces cisplatin resistance through upregulation of xCT expression and glutathione synthesis in gastric cancer

doi: 10.1111/cas.15869

Figure Lengend Snippet: Proposed mechanism of growth differentiation factor 15 (GDF15)‐mediated cisplatin resistance. In the present study, we found that GDF15‐elevated glutathione (GSH) through the glial cell‐derived neurotrophic factor family receptor a‐like (GFRAL)‐general control nonderepressible 2 (GCN2)‐eukaryotic initiation factor 2α (eIF2α)‐activating transcription factor 4 (ATF4)‐xCT pathway enhances cisplatin resistance for gastric cancer. Figure was created by Servier Medical Art. PKR, protein kinase R; RET, rearranged during transfection; ROS, reactive oxygen species.

Article Snippet: The GDF15 neutralizing Ab (MAB957) and recombinant human GDF15 ( Escherichia coli ‐expressed) protein (#9279‐GD‐050) were obtained from R&D Systems.

Techniques: Derivative Assay, Control, Transfection