Journal: Nature

Article Title: PTER is a N -acetyltaurine hydrolase that regulates feeding and obesity

doi: 10.1038/s41586-024-07801-6

Figure Lengend Snippet: a , b , Change in body weight ( a ) and food intake ( b ) of 26–28-week-old male DIO C57BL/6J mice following 7 days of treatment with the indicated dose of N -acetyltaurine (NAT; i.p.). For saline versus N -acetyltaurine (15 mg per kg per day), P = 5.95 × 10 –4 ; for saline versus N -acetyltaurine (50 mg per kg per day), P = 6.3 × 10 –4 . N = 5 per group for vehicle, 1 and 5 mg per kg per day; N = 6 per group for 15 and 50 mg per kg per day. c , d , Change in body weight ( c ) and food intake ( d ) of 19–21-week-old male DIO C57BL/6J mice following treatment with the indicated metabolite at a dose of 15 mg per kg per day (i.p.). N = 5 per group. e – g , Western blots with anti-PTER (top) and anti-tubulin (bottom) antibodies ( e ), N -acetyltaurine hydrolysis activity ( f ) and tissue N -acetyltaurine levels ( g ) from cortex (Cort.), hypothalamus (Hyp.) and brainstem (BS) of WT mice and Pter KO mice. For WT versus Pter KO brainstem, P = 6.65 × 10 –4 . N = 6 per group for f and g . h , Change in 24-h food intake of 6-month-old male DIO mice treated with a single dose of GDF15 (0.1 mg kg –1 , i.p.) in the presence of anti-GFRAL antibody (10 mg kg –1 , i.p.) or IgG control antibody (10 mg kg –1 , i.p.). N = 5 per group. i , j , Change in body weight ( i ) and cumulative food intake ( j ) of 16-week-old male DIO mice following saline or N -acetyltaurine (15 mg per kg per day, i.p.) treatment and with IgG or anti-GFRAL antibody co-treatment (10 mg kg –1 , i.p., once every 3 days). N = 10 per group. Data are shown as the mean ± s.e.m. For e , the loading control was performed on the same blot. In a – d and f – h , P values were calculated from two-tailed unpaired t -tests and were not corrected for multiple comparisons. In i and j , P values were calculated from two-way ANOVA with post hoc Sidak’s multiple comparisons test. All experiments were performed once.

Article Snippet: Recombinant GDF15 (957-GD) was purchased from R&D Systems.

Techniques: Saline, Western Blot, Activity Assay, Control, Two Tailed Test

Journal: Nature

Article Title: PTER is a N -acetyltaurine hydrolase that regulates feeding and obesity

doi: 10.1038/s41586-024-07801-6

Figure Lengend Snippet: Related to Fig. . a - d , Body weight ( a , b ) and food intake ( c , d ) of 6 to 7-month-old DIO male C57BL/6J mice after a 7-day treatment of saline or N-acetyltaurine (15 mg/kg/day, IP) or GLP-1 (2 mg/kg/day, IP) with or without Exendin-3 (0.1 mg/kg/day, IP). N = 7/group. NAT, N-acetyltaurine. e , f , Body weight ( e ) and food intake ( f ) of 5-month-old DIO male C57BL/6J mice or 3 to 4-month-old MC4R-KO mice on high fat diet after a 7-day treatment of saline or N-acetyltaurine (15 mg/kg/day, IP). N = 6/group. NAT, N-acetyltaurine. g , Plasma GDF15 (left), GLP-1 (middle) and leptin (right) levels of 19 to 21-week-old male DIO C57BL/6J mice following treatment with N-acetyltaurine 15 mg/kg/day (IP) or saline. N = 5 per group. NAT, N-acetyltaurine. Data are shown as mean ± SEM. P-values were calculated from two-tailed unpaired t-tests.

Article Snippet: Recombinant GDF15 (957-GD) was purchased from R&D Systems.

Techniques: Saline, Clinical Proteomics, Two Tailed Test

Journal: Clinical and Experimental Medicine

Article Title: GDF15 activates human fibroblast MRC5 cells via miR-338/STAT1 in silicosis

doi: 10.1007/s10238-025-01627-w

Figure Lengend Snippet: Increased blood GDF15 expression in patients with silicosis. A , Serum GDF15 expression in patients with silicosis was higher than that in controls ( P = 0.0003, compared to control). B , Increased serum GDF15 expression did not relate to the FEV1% of patients with silicosis; P = 0.2002. C , Increased serum GDF15 expression did not relate to the clinical stages of silicosis, P = 0.5979

Article Snippet: Recombinant human GDF15 was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Control

Journal: Clinical and Experimental Medicine

Article Title: GDF15 activates human fibroblast MRC5 cells via miR-338/STAT1 in silicosis

doi: 10.1007/s10238-025-01627-w

Figure Lengend Snippet: GDF15 activates human embryonic lung fibroblast MRC5 cells. The effect of GDF15 on col1a and α-SMA protein expression in MRC5 cells was detected using western blotting. A , dosage course. B , time course. C , GDF15 increased the proliferation of MRC cells, as detected in the CCK-8 assay. D, GDF15 increased the migration of MRC cells, which was detected in the wound healing assay. Data represent the mean ± SD from three independent experiments.*,**, ♯ , P < 0.05

Article Snippet: Recombinant human GDF15 was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Western Blot, CCK-8 Assay, Migration, Wound Healing Assay

Journal: Clinical and Experimental Medicine

Article Title: GDF15 activates human fibroblast MRC5 cells via miR-338/STAT1 in silicosis

doi: 10.1007/s10238-025-01627-w

Figure Lengend Snippet: Effects of GDF15 on the miRNA and mRNA expression in MRC5 cells. Effect of GDF15 on the miRNA profile of MRC5 cells. A , heatmap; B , Volcano plots; C , KEGG analysis. Effect of GDF15 on the mRNA profile of MRC5 cells. D , heatmap; E, Volcano plots; F, KEGG analysis

Article Snippet: Recombinant human GDF15 was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing

Journal: Clinical and Experimental Medicine

Article Title: GDF15 activates human fibroblast MRC5 cells via miR-338/STAT1 in silicosis

doi: 10.1007/s10238-025-01627-w

Figure Lengend Snippet: GDF15 reduced the expression of miR-338 in MRC5 cells. A , qRT-PCR results showed that the expression of miR-338 was decreased in response to GDF15 expression. B , Western blotting results showed that miR-338 mimics attenuated the expression of col1a and α-SMA regardless of treatment with GDF15, whereas miR-338 inhibitors promoted the expression of col1 and α-SMA regardless of treatment with GDF15. C , CCK-8 assay results showed that miR-338 mimics reduced the growth rate and the miR-338 inhibitor increased the growth rate of MRC5 cells. Data represent the mean ± SD from three independent experiments.*,**, ♯ , P < 0.05

Article Snippet: Recombinant human GDF15 was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, CCK-8 Assay

Journal: Clinical and Experimental Medicine

Article Title: GDF15 activates human fibroblast MRC5 cells via miR-338/STAT1 in silicosis

doi: 10.1007/s10238-025-01627-w

Figure Lengend Snippet: GDF15 increased STAT1 expression in MRC5 cells. A , Western blotting results showed that GDF15 induced STAT1 expression. B , Western blotting results showed that STAT1 siRNA attenuated the expression of col1a and α-SMA, whereas STAT1 cDNA promoted their expression in the presence or absence of GDF15 treatment. C , CCK-8 assay results showed that STAT1 siRNA reduced the growth rate, whereas STAT1 cDNA increased the growth rate of MRC5 cells. Data represent the mean ± SEM from three independent experiments.*,**, ♯ , P < 0.05

Article Snippet: Recombinant human GDF15 was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Expressing, Western Blot, CCK-8 Assay

Journal: Clinical and Experimental Medicine

Article Title: GDF15 activates human fibroblast MRC5 cells via miR-338/STAT1 in silicosis

doi: 10.1007/s10238-025-01627-w

Figure Lengend Snippet: GDF15 activated MRC5 cells through the miR-338/STAT1 pathway. A , Using TargetScan ( http://www.targetscan.org/ ), the conserved miR-338 binding site in the 3’-UTR of STAT1 mRNA was constructed in the pmirGLO dual-luciferase miRNA target expression vector. Luciferase activity was analyzed in the MRC5 cells. MRC5 cells were co-transfected with miR-30 mimics and a luciferase reporter. B , miR-338 regulated STAT1 protein expression. C , miR-338 mimics attenuated the expression of col1 and α-SMA protein induced by STAT1 overexpression upon co-treatment with GDF15. D, STAT1 knockdown attenuated col1 and α-SMA protein expression induced by the miR-338 inhibitor upon co-treatment with GDF15. Data represent the means ± SD from three independent experiments. *,**, ♯ , ♯♯ , P < 0.05

Article Snippet: Recombinant human GDF15 was purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Binding Assay, Construct, Luciferase, Expressing, Plasmid Preparation, Activity Assay, Transfection, Over Expression, Knockdown

Journal: Journal for immunotherapy of cancer

Article Title: Loss of ADAR1 in macrophages in combination with interferon gamma suppresses tumor growth by remodeling the tumor microenvironment.

doi: 10.1136/jitc-2023-007402

Figure Lengend Snippet: Figure 3 ADAR1 loss in macrophage with IFN-γ treatment affects the secretion of key cytokines through PKR/EIF2α signaling. (A, C) Human XL cytokine arrays for detecting differential factors between THP-1 cells with scrambled shRNA and shADAR1#1 under the treatment of IFN-γ (A) and between THP-1 cells with empty vector and WT ADAR, co-cultured with A549 (C). (B, D) Bar plots showing the expression levels of differential factors on ADAR1 knockdown (B) and ADAR1 overexpression (D). (E) Venn gram showing key cytokines identified by ADAR1 knockdown and overexpression experiments and their potential effects on the tumor microenvironment. (F) RT-qPCR-based mRNA expression levels of ADAR, CCL20, GDF15, IFN-G, IL-18, IL-18BP, and HAVCR2 in different THP-1 cells (transfected with scrambled shRNA, shADAR1#1 or shADAR1#2) with IFN-γ treatment. β-actin was used as an internal control. (G) RT-qPCR-based mRNA expression levels of Ccl20, Gdf15, Il-18, Il-18bp, and Havcr2 in BMDMs from C57BL/6 mice (Adarfl/fl and Adarfl/flLyz2Cre) treated with IFN-γ. Gapdh was used as an internal control. (H) Immunofluorescent staining for anti-dsRNA (J2) in THP-1 cells treated with IFN-γ. RNase III treatment was used as the negative control for the dsRNA signal. Scale bars, 10 µm. (I) Western blot showing the protein expression of p-PKRThr446/

Article Snippet: 2- AP (GlpBio, GC61906), Phorbol 12- myristate 13- acetate (PMA) (MedChemExpress, HY- 18739), Recombinant Human IFN-γ (Novoprotein, C014), Recombinant Mouse IL- 2 (Novoprotein, P04351), Recombinant M- CSF (Novoprotein, CB34), Recombinant Mouse IFN-γ (Novoprotein, C746), Recombinant Mouse TIM- 3 (Novoprotein, CM54), Recombinant Mouse CCL20 (PeproTech, 250–2), Recombinant Mouse GDF15 (R&D Systems, 8944- GD025), Recombinant Mouse IL- 18 Binding Protein Isoform d (Novoprotein, CM45), ShortCut RNase III (NEB, M0245S), poly (I:C) (GlpBio, GC14710).

Techniques: shRNA, Plasmid Preparation, Cell Culture, Expressing, Knockdown, Over Expression, Quantitative RT-PCR, Transfection, Control, Staining, Negative Control, Western Blot

Journal: Journal for immunotherapy of cancer

Article Title: Loss of ADAR1 in macrophages in combination with interferon gamma suppresses tumor growth by remodeling the tumor microenvironment.

doi: 10.1136/jitc-2023-007402

Figure Lengend Snippet: Figure 5 The effect of ADAR1 loss and key cytokines on tube formation. (A) Schematic diagram of tube formation experiment of SVEC4-10 cells. (B) Tube formation of SVEC4-10 cells treated with conditioned media from BMDMs (Adarfl/fl vs Adarfl/flLyz2Cre) and CCL20, GDF15, or IFN-γ. Scale bar, 500 µm. (C) Bar plots showing relative vessel area, the total number of junctions, and the vessel length for SVEC4-10 tube formation. (D) Tube formation of SVEC4-10 cells treated with conditioned media from BMDMs (Adarfl/flLyz2Cre) and IFN-γ after pretreatment with or without 2-AP (5 mM). Scale bar, 500 µm. (E) Bar plots showing relative vessel area, the total number of junctions, and vessel length for SVEC4-10 tube formation. (F) Tube formation of HUVEC cells treated with conditioned media from THP-1 cells (transfected with shADAR1#1) and IFN-γ after pretreatment with or without 2-AP (5 mM). Scale bar, 200 µm. (G) Bar plots showing relative vessel area, the total number of junctions, and the vessel length for HUVEC tube formation. (C, E, and G) P values are based on unpaired Student’s t-test. Data are presented as mean±SD. ADAR, adenosine deaminases acting on RNA; BMDMs, bone marrow-derived macrophages; IFN, interferon; 2-AP, 2-aminopurine.

Article Snippet: 2- AP (GlpBio, GC61906), Phorbol 12- myristate 13- acetate (PMA) (MedChemExpress, HY- 18739), Recombinant Human IFN-γ (Novoprotein, C014), Recombinant Mouse IL- 2 (Novoprotein, P04351), Recombinant M- CSF (Novoprotein, CB34), Recombinant Mouse IFN-γ (Novoprotein, C746), Recombinant Mouse TIM- 3 (Novoprotein, CM54), Recombinant Mouse CCL20 (PeproTech, 250–2), Recombinant Mouse GDF15 (R&D Systems, 8944- GD025), Recombinant Mouse IL- 18 Binding Protein Isoform d (Novoprotein, CM45), ShortCut RNase III (NEB, M0245S), poly (I:C) (GlpBio, GC14710).

Techniques: Transfection, Derivative Assay

Journal: Journal for immunotherapy of cancer

Article Title: Loss of ADAR1 in macrophages in combination with interferon gamma suppresses tumor growth by remodeling the tumor microenvironment.

doi: 10.1136/jitc-2023-007402

Figure Lengend Snippet: Figure 8 Schematic summary of this study. ADAR1-deficient macrophages combined with IFN-γ treatment reprogram the tumor microenvironment by two mechanisms: (1) inhibit angiogenesis by decreased secretion of GDF15 and CCL20 and increased secretion of IFN-γ, and (2) activate CD8+ T cells by decreased secretion of TIM-3 and IL-18BP and increased secretion of IL-18. These effects collectively convert a “cold tumor” into a “hot tumor.” Combined treatment with ADAR1- deficient macrophages and IFN-γ may represent an effective therapeutic approach. dsRNA, double-stranded RNA; IFN, interferon; IL, interleukin; uORF, upstream Open Reading Frame.

Article Snippet: 2- AP (GlpBio, GC61906), Phorbol 12- myristate 13- acetate (PMA) (MedChemExpress, HY- 18739), Recombinant Human IFN-γ (Novoprotein, C014), Recombinant Mouse IL- 2 (Novoprotein, P04351), Recombinant M- CSF (Novoprotein, CB34), Recombinant Mouse IFN-γ (Novoprotein, C746), Recombinant Mouse TIM- 3 (Novoprotein, CM54), Recombinant Mouse CCL20 (PeproTech, 250–2), Recombinant Mouse GDF15 (R&D Systems, 8944- GD025), Recombinant Mouse IL- 18 Binding Protein Isoform d (Novoprotein, CM45), ShortCut RNase III (NEB, M0245S), poly (I:C) (GlpBio, GC14710).

Techniques:

Journal: Oncotarget

Article Title: GDF15 contributes to radioresistance and cancer stemness of head and neck cancer by regulating cellular reactive oxygen species via a SMAD-associated signaling pathway

doi: 10.18632/oncotarget.13649

Figure Lengend Snippet: A. Administration of rhGDF15 led to radioresistance in two HNC cell lines. Detroit and KB cells were treated with 20 ng/ml rhGDF15 for 5 days and then subjected to various doses of irradiation (0, 2, and 4 Gy). Fourteen days later, the cell colonies were determined by staining with 0.05% crystal violet (n=3). B. GDF15 expression was successfully inhibited after transfection of GDF15-specific shRNA plasmids (GDF15sh) in KB and OECM1 cell lines. After 48 h, cells were harvested for western blot analysis. C. Silencing GDF15 sensitized cells to irradiation in HNC cells. OECM1 and KB cells were transfected with GDF15sh or the vector plasmids and then subjected to irradiation (0 to 6 Gy). After 14 days, the cell colonies were determined (n=3). D. GDF15 had minimal effect on growth regulation in HNC cells. The OECM1, Detroit and KB cells were treated with or without rhGDF15 (20 ng/ml for 5 days). After 14 days, the cell colonies were determined (n=3). (*: p <0.05, n.s. : non-significance, t -test).

Article Snippet: For administration of GDF15, the recombinant human GDF15 protein (rhGDF15) (#957-GD, R&D Systems, Minneapolis, MN) was added to the cell culture medium at 20 ng/ml for 2 to 5 days.

Techniques: Irradiation, Staining, Expressing, Transfection, shRNA, Western Blot, Plasmid Preparation

Journal: Oncotarget

Article Title: GDF15 contributes to radioresistance and cancer stemness of head and neck cancer by regulating cellular reactive oxygen species via a SMAD-associated signaling pathway

doi: 10.18632/oncotarget.13649

Figure Lengend Snippet: A. Administration of rhGDF15 suppressed intracellular ROS level in HNC cells. KB and OECM1 cells were treated with 20 ng/ml of rhGDF15 for 5 days. After staining with DCF dye, the cells were subjected to flow cytometric analysis to determine the intracellular ROS levels. All values are presented as fluorescence intensity. B. GDF15 silencing increased ROS production in HNC cells. After transfection with GDF15sh or the vector plasmids, the KB and OECM1 cells were stained with DCF dye and subjected to flow cytometric analysis. C. GDF15 silencing increased ROS production in esophageal cancer cells. CE48T/VGH and CE81T/VGH cells were co-transfected with DsRed and GDF15sh/vector plasmids and then subjected to 4 Gy of irradiation. After staining with DCF dye, the cells were subjected to confocal microscopy. D. GDF15 silencing reduced mitochondrial membrane potential and led to apoptosis in HNC cells. KB or Detroit cells were transfected with the GDF15sh or the vector plasmids for 48 h. After incubating with MitoCapture reagents, the cells were subjected to flow cytometry analysis. The PE fluorescence fraction represents healthy cells, and the FITC fraction represents apoptotic cells.

Article Snippet: For administration of GDF15, the recombinant human GDF15 protein (rhGDF15) (#957-GD, R&D Systems, Minneapolis, MN) was added to the cell culture medium at 20 ng/ml for 2 to 5 days.

Techniques: Staining, Fluorescence, Transfection, Plasmid Preparation, Irradiation, Confocal Microscopy, Membrane, Flow Cytometry

Journal: Oncotarget

Article Title: GDF15 contributes to radioresistance and cancer stemness of head and neck cancer by regulating cellular reactive oxygen species via a SMAD-associated signaling pathway

doi: 10.18632/oncotarget.13649

Figure Lengend Snippet: A. Elevation of GDF15 mRNA expressions in CD44+ sorted cells compared to the CD44- cells in both KB and OECM1 cell lines, as determined by RT-PCR method (n=3). B. Elevation of GDF15 protein levels in the conditional medium of CD44+ sorted cells compared to the CD44- cells in both KB and OECM1 cell lines, as determined by ELISA method (n=3). C. Elevation of GDF15 mRNA expressions in ALDH1+ sorted cells compared to the ALDH1- cells in both KB and OECM1 cell lines, as determined by RT-PCR method (n=3). D. Elevation of CD44 mRNA expressions in ALDH1+ sorted cells compared to the ALDH1- cells in both KB and OECM1 cell lines, as determined by RT-PCR method (n=3). E. Elevation of CD44+ population in rhGDF15 treated cells compared to non-treated cells in both KB and Detroit cell lines, as determined by FACS method. F. Reduction of CD44+ population in GDF15sh transfected cells in both KB and Detroit cell lines, as determined by FACS method. G. Elevation of ALDH1+ population in rhGDF15 treated cells in both KB and Detroit cell lines, as determined by FACS method. H. Reduction of ALDH1+ population in GDF15sh transfected cells in both KB and Detroit cell lines, as determined by FACS method. I. Enhancement of spheroid cell formation in the GDF15+ sorted cells compared to GDF15- cells in both KB and OECM1 cell lines (n=3). J. Reduction of spheroid cell formation in GDF15sh transfected cells in both Detroit and KB cell lines (n=3). K. Increase of invasion ability in the rhGDF15 treated cells compared to non-treated cells (control) in both Detroit and KB cell lines (n=3). L. Increase of migration ability in the rhGDF15 treated cells compared to non-treated cells (control) in both Detroit and KB cell lines (n=3). (*: p <0.05, **: p <0.01, ***: p <0.001, n.s. : non-significance, t -test).

Article Snippet: For administration of GDF15, the recombinant human GDF15 protein (rhGDF15) (#957-GD, R&D Systems, Minneapolis, MN) was added to the cell culture medium at 20 ng/ml for 2 to 5 days.

Techniques: Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Transfection, Control, Migration

Journal: Oncotarget

Article Title: GDF15 contributes to radioresistance and cancer stemness of head and neck cancer by regulating cellular reactive oxygen species via a SMAD-associated signaling pathway

doi: 10.18632/oncotarget.13649

Figure Lengend Snippet: A. Treatment of antioxidant agent led to radioresistance in HNC cells. After treatment of 10 μM N-acetylcysteine (NAC) for 48 h, Detroit or KB cells were subjected to serial dose of irradiation (0 to 6 Gy). Fourteen days later, the numbers of surviving cell colonies were determined (n=3). B. Treatment of antioxidant agent increase CD44+ cell population. After treatment of 10 μM NAC for 48 h, the OECM1 or KB cells were subjected to flow cytometry analysis for CD44+ populations. C. GDF15 reversed the effects of ROS in the suppression of spheroid cell formation. Detroit or KB cells were treated with H 2 O 2 (5 μM) with or without addition of rhGDF15 (20 ng/ml) for 48 h. These cells were then incubated in the spheroid cell culture condition and assessed after 14 days (n=3). (*: p < 0.05, **: p < 0.01, ***: p < 0.001, t -test).

Article Snippet: For administration of GDF15, the recombinant human GDF15 protein (rhGDF15) (#957-GD, R&D Systems, Minneapolis, MN) was added to the cell culture medium at 20 ng/ml for 2 to 5 days.

Techniques: Irradiation, Flow Cytometry, Incubation, Cell Culture

Journal: Oncotarget

Article Title: GDF15 contributes to radioresistance and cancer stemness of head and neck cancer by regulating cellular reactive oxygen species via a SMAD-associated signaling pathway

doi: 10.18632/oncotarget.13649

Figure Lengend Snippet: A. GDF15 expression was reduced after treatment of TGF-β inhibitor. KB and OECM1 cells were treated with serial doses of LY364947 (5 to 20 μM) for 24 h. Cellular protein was extracted and subjected to western blot analysis to assess GDF15 protein expression. B. Administration of rhGDF15 increased TGF-β downstream molecule PAI-1 luciferase reporter activity. KB or Detroit cells were transfected with luciferase reporter plasmid carrying PAI-1 gene, with or without addition of rhGDF15 (20 ng/ml). After 48 h, cells were harvested for measurements of luciferase activity (n=3). C. Silencing rhGDF15 suppressed PAI-1 luciferase reporter activity. KB or Detroit cells were transfected with luciferase reporter plasmid carrying PAI-1 gene, with or without co-transfection of GDF15sh plasmid. After 48 h, cells were harvested for measurements of luciferase activity (n=3). D. PAI-1 expression induced by TGF-β was suppressed following GDF15 silencing. KB or Detroit cells were transfected with luciferase reporter plasmid carrying PAI-1 gene, with or without addition of rhGDF15 (20 ng/ml), or with or without co-transfection of GDF15sh plasmid. After 48 h, cells were harvested for measurements of luciferase activity (n=3). (*: p < 0.05, **: p < 0.01, ***: p < 0.001, t -test).

Article Snippet: For administration of GDF15, the recombinant human GDF15 protein (rhGDF15) (#957-GD, R&D Systems, Minneapolis, MN) was added to the cell culture medium at 20 ng/ml for 2 to 5 days.

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

Journal: Oncotarget

Article Title: GDF15 contributes to radioresistance and cancer stemness of head and neck cancer by regulating cellular reactive oxygen species via a SMAD-associated signaling pathway

doi: 10.18632/oncotarget.13649

Figure Lengend Snippet: A, B. Administration of rhGDF15 increased the phosphorylated forms of SMAD family proteins in HNC cells. KB or FaDu cells were treated with serial doses of rhGDF15 (0-10 ng/ml) for 15 min (A) or 5 ng/ml rhGDF15 for various times (0-120 min) (n=3). (B) The cellular proteins were extracted and subjected to western blot analysis for SMAD family protein expressions (n=3). C. Effects of SMAD1-siRNA on the expressions of SMAD family proteins. After transfection of SMAD1-specific siRNA or the scramble oligonucleotides in HNC cells for 48h, cellular proteins were extracted for western blot analysis. GAPDH protein was used as an internal control (n=3). D. Silencing SMAD1 increased ROS level in HNC cells. KB or OECM1 cells were transfected SMAD1 specific siRNA or the scramble oligonucleotides for 48 h. After treating cells with 10 μM H 2 O 2 for 20 min, the Intracellular ROS levels were determined by DCF dye staining and analyzed with flow cytometry. E. Silencing SMAD1 suppressed spheroid cell formation in HNC cells. Fadu, OECM1 or KB cells were transfected SMAD1 specific siRNA or the scramble oligonucleotides for 48 h. These cells were then incubated in the spheroid cell culture condition and assessed after 14 days (n=3). F. The spheroid cell formation promoted by GDF15 was inhibited in SMAD knockdown HNC cells. KB or OECM1 cells were transfected SMAD1 specific siRNA or the scramble oligonucleotides for 48 h, with the addition of rhGDF15 (20 ng/ml). These cells were then incubated in the spheroid cell culture condition and assessed after 14 days (n=3). G. Effects of SMAD3-siRNA on the expressions of SMAD family proteins. After transfection of SMAD3-specific siRNA or the scramble oligonucleotides in HNC cells for 48h, cellular proteins were extracted for western blot analysis. GAPDH protein was used as an internal control (n=3). H. Silencing SMAD3 had no effect on ROS level in HNC cells. Fadu or OECM1 cells were transfected SMAD3-specific siRNA or the scramble oligonucleotides for 48 h. After treating cells with 10 μM H 2 O 2 for 20 min, the Intracellular ROS levels were determined by DCF dye staining and analyzed with flow cytometry. I. Silencing SMAD3 had no significant effect on spheroid cell formation in HNC cells. Fadu, OECM1 or Detroit cells were transfected SMAD3 specific siRNA or the scramble oligonucleotides for 48 h, with the addition of rhGDF15 (20 ng/ml). After 14 days of incubation in the spheroid cell culture condition, cells were assessed for spheroid formation (n=3). (*: p < 0.05, **: p < 0.01, ***: p < 0.001, n.s. : non-significance, t -test).

Article Snippet: For administration of GDF15, the recombinant human GDF15 protein (rhGDF15) (#957-GD, R&D Systems, Minneapolis, MN) was added to the cell culture medium at 20 ng/ml for 2 to 5 days.

Techniques: Western Blot, Transfection, Control, Staining, Flow Cytometry, Incubation, Cell Culture, Knockdown

Journal: Oncotarget

Article Title: GDF15 contributes to radioresistance and cancer stemness of head and neck cancer by regulating cellular reactive oxygen species via a SMAD-associated signaling pathway

doi: 10.18632/oncotarget.13649

Figure Lengend Snippet: A total of 4×10 6 KB cells, with or without pre-treatment with the rhGDF15 protein (20 ng/ml for 5 days), were subcutaneously injected into BALB/c mice (10 mice each group) in the upper portion of the hind limb. At day 14, each group was randomly divided into two groups (5 mice per group), with or without receiving 2 Gy of irradiation, followed by repeated irradiation of the same dose twice a week for a total of 8 Gy. A. Tumor volume was measured twice a week and calculated as (length x width x height) for 36 days. B-D. The tumors in the group of irradiation, either with or without pre-treatment of rhGDF15, were dissected. The protein expression levels of SMAD family molecules in the rhGDF15 treatment tumor group (B) or the control groups (C) were determined by using western blot analysis, and quantified the relative expression levels after normalized with GAPDH (D). E. The expression levels of ALDH1 and Nestin in tumor tissues were determined by using IHC analysis. Three tumor sections of IHC staining were shown for examples (*: p < 0.05, **: p < 0.01, ***: p < 0.001, t -test).

Article Snippet: For administration of GDF15, the recombinant human GDF15 protein (rhGDF15) (#957-GD, R&D Systems, Minneapolis, MN) was added to the cell culture medium at 20 ng/ml for 2 to 5 days.

Techniques: Injection, Irradiation, Expressing, Control, Western Blot, Immunohistochemistry

Journal: Oncotarget

Article Title: GDF15 contributes to radioresistance and cancer stemness of head and neck cancer by regulating cellular reactive oxygen species via a SMAD-associated signaling pathway

doi: 10.18632/oncotarget.13649

Figure Lengend Snippet: Diagram of the mechanism by which GDF15 contributes to radioresistance and cancer stemness through regulating ROS levels via a SMAD-associated pathway

Article Snippet: For administration of GDF15, the recombinant human GDF15 protein (rhGDF15) (#957-GD, R&D Systems, Minneapolis, MN) was added to the cell culture medium at 20 ng/ml for 2 to 5 days.

Techniques:

Journal: Journal of cachexia, sarcopenia and muscle

Article Title: GDNF family receptor alpha-like antagonist antibody alleviates chemotherapy-induced cachexia in melanoma-bearing mice.

doi: 10.1002/jcsm.13219

Figure Lengend Snippet: Figure 1 Anti-GFRAL antibodies were selected by solution-phase biopanning. (A) Strategy for anti-GFRAL antibody selection using a human combina- torial antibody phage library. Black dotted lines represent GFRAL-binding phage pool. (B) Polyclonal phage ELISA for phage pools from each round of biopanning. (C) Heavy-chain complementarity-determining region 3 (H-CDR3) amino acid sequences of the anti-GFRAL antibody clones. (D) Binding of the anti-GFRAL antibodies to both human and mouse GFRAL extracellular domains verified by ELISA. (E) Luciferase reporter assay of HEK293 cells transfected with human GFRAL, human RET, and SRE-luciferase genes for the selection of the most potent GFRAL antagonist antibody with inhibitory activity against GDF15-induced luminescent signal (n = 5). Data are presented as the mean ± standard error of the mean (SEM), analysed with the Kruskal–Wallis test, followed by the uncorrected Dunn’s test. Statistical differences in post hoc testing are indicated as ns = non-significant, ***P < 0.005.

Article Snippet: Following that, 5-fold serial dilutions of A11 starting at 20 μg/mL or unbiotinylated recombinant human GDF15 (8146-GD; R&D Systems) controls starting at 180 μM were added to plates for 1 h at 37°C.

Techniques: Selection, Binding Assay, Enzyme-linked Immunosorbent Assay, Clone Assay, Luciferase, Reporter Assay, Transfection, Activity Assay

Journal: Journal of cachexia, sarcopenia and muscle

Article Title: GDNF family receptor alpha-like antagonist antibody alleviates chemotherapy-induced cachexia in melanoma-bearing mice.

doi: 10.1002/jcsm.13219

Figure Lengend Snippet: Figure 2 The antibody A11 binds to GFRAL and inhibits the signal transduction of the GDF15/GFRAL/RET axis. (A) SPR sensorgram showing the binding kinetics of the antibody A11 to the recombinant human GFRAL extracellular domain. Black box indicates concentration of antibody A11 (1.89– 31.25 nM). (B) Flow cytometry analysis of the control antibody and antibody A11 binding in wild-type (WT) and human GFRAL gene-transfected HEK293 cells. Black box indicates concentration of antibody A11 (0–50 nM). (C) Representative immunostaining images of commercial anti-GFRAL an- tibody (left) and antibody A11 (right) binding in WT and human GFRAL gene-transfected HEK293 cells. The white scale bar represents 30 μm. (D) West- ern blot analysis showing the inhibitory effect of the antibody A11 on the phosphorylation of RET, AKT, and ERK in human GFRAL/RET gene-transfected HEK293 cells. The gel is representative of three independent experiments.

Article Snippet: Following that, 5-fold serial dilutions of A11 starting at 20 μg/mL or unbiotinylated recombinant human GDF15 (8146-GD; R&D Systems) controls starting at 180 μM were added to plates for 1 h at 37°C.

Techniques: Transduction, Binding Assay, Recombinant, Concentration Assay, Flow Cytometry, Control, Transfection, Immunostaining, Phospho-proteomics

Journal: Journal of cachexia, sarcopenia and muscle

Article Title: GDNF family receptor alpha-like antagonist antibody alleviates chemotherapy-induced cachexia in melanoma-bearing mice.

doi: 10.1002/jcsm.13219

Figure Lengend Snippet: Figure 3 The antibody A11 attenuates cisplatin-induced cachexia in vivo Normal: control mice with PBS injections, Cis/Con: mice with cisplatin and control antibody injections, Cis/A11: mice with cisplatin and the antibody A11 injections (n = 7 mice per group). (A) Cumulative food intake per mouse on days 2–5. (B) Body weight change compared with the weight before drug administration followed up to day 5. (C) Weights of isolated skeletal mus- cles (quadriceps, gastrocnemius, and soleus). (D) Weights of isolated adipose tissues (eWAT and iWAT). (E) Plasma levels of GDF15 on day 5, deter- mined by ELISA. Data are presented as the mean ± SEM, analysed with the Kruskal–Wallis test, followed by the uncorrected Dunn’s test. Statistical differences in post hoc testing are indicated as ns = non-significant, *P < 0.05, **P < 0.01, ***P < 0.005, and ****P < 0.001.

Article Snippet: Following that, 5-fold serial dilutions of A11 starting at 20 μg/mL or unbiotinylated recombinant human GDF15 (8146-GD; R&D Systems) controls starting at 180 μM were added to plates for 1 h at 37°C.

Techniques: In Vivo, Control, Isolation, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Journal of cachexia, sarcopenia and muscle

Article Title: GDNF family receptor alpha-like antagonist antibody alleviates chemotherapy-induced cachexia in melanoma-bearing mice.

doi: 10.1002/jcsm.13219

Figure Lengend Snippet: Figure 4 The antibody A11 suppresses c-Fos accumulation on the hindbrain of cisplatin-treated mice Normal: control mice with PBS injections, Cis/ Con: mice with cisplatin and control antibody injections, Cis/A11: mice with cisplatin and the antibody A11 injections (n = 5 mice per group). (A) Rep- resentative sections of the area postrema (AP) and the nucleus of the solitary tract (NTS) showing c-Fos expression in GFRAL-expressing neurons 4 h after cisplatin treatment on day 4. White arrows represent neurons stained with both GFRAL and c-Fos, indicating activation by cisplatin-induced GDF15. The white scale bars represent 150 μm and the yellow scale bar represents 20 μm. The CC represents central canal. (B) Quantification of cisplatin-induced c-Fos expression in the AP/NTS 4 h after cisplatin injection. Each point represents one mouse. Data from each mouse represent quan- tification from 6 alternative sections. (C) Number of GFRAL-positive neurons in the AP/NTS that co-express c-Fos 4 h after cisplatin injection. Each point represents one mouse. Data from each mouse represent quantification from 6 alternative sections. Data are presented as the mean ± SEM, analysed with the Kruskal–Wallis test, followed by the uncorrected Dunn’s test. Statistical differences in post hoc testing are indicated as ns = non-significant, *P < 0.05, **P < 0.01.

Article Snippet: Following that, 5-fold serial dilutions of A11 starting at 20 μg/mL or unbiotinylated recombinant human GDF15 (8146-GD; R&D Systems) controls starting at 180 μM were added to plates for 1 h at 37°C.

Techniques: Control, Expressing, Staining, Activation Assay, Injection

Journal: Journal of cachexia, sarcopenia and muscle

Article Title: GDNF family receptor alpha-like antagonist antibody alleviates chemotherapy-induced cachexia in melanoma-bearing mice.

doi: 10.1002/jcsm.13219

Figure Lengend Snippet: Figure 5 The antibody A11 ameliorates cisplatin-induced cachexia in a mouse model of melanoma Normal: control mice without tumour, TB/Con: tu- mour-bearing mice with control antibody injections, TB/A11: tumour-bearing mice with the antibody A11 injections, TB/Cis/Con: tumour-bearing mice with cisplatin and control antibody injections, TB/Cis/A11: tumour-bearing mice with cisplatin and the antibody A11 injections (n = 10–16 mice per group). (A) Cumulative food intake per mouse on days 13–18. (B) Body weight change compared with the weight before drug administration followed up to day 17. (C) Weights of isolated skeletal muscles (quadriceps, gastrocnemius, and soleus). (D) Weights of isolated adipose tissues (eWAT and iWAT). (E) Plasma GDF15 levels on day 18, determined by ELISA. Data are presented as the mean ± SEM, analysed with the Kruskal–Wallis test, followed by the uncorrected Dunn’s test. Statistical differences in post hoc testing are indicated as ns = non-significant, *P < 0.05, **P < 0.01, ***P < 0.005, and ****P < 0.001.

Article Snippet: Following that, 5-fold serial dilutions of A11 starting at 20 μg/mL or unbiotinylated recombinant human GDF15 (8146-GD; R&D Systems) controls starting at 180 μM were added to plates for 1 h at 37°C.

Techniques: Control, Isolation, Muscles, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Identification of a distinct cluster of GDF15 high macrophages exhibiting anti-inflammatory activities

doi: 10.1101/2024.01.11.575315

Figure Lengend Snippet: Immunofluorescence double labeling showing the existence of GDF15 high macrophages in lung tissues. (A) Results obtained in healthy human lung tissues from 3 independent subjects. Two representative microscopic fields were shown. Arrowheads indicated the CD68 + GDF15 high macrophages. (B) Results obtained in human lung tissues with COPD from 4 independent subjects. Arrowheads indicated the CD68 + GDF15 high macrophages. (C) Results obtained in rat lung tissues without and with experimental PAH. The nuclei were counterstained with DAPI (blue). White arrowheads indicated CD68 + GDF15 high macrophages. The red box highlighted the presence of CD68 + GDF15 high macrophages (white arrowheads); the green box highlighted the presence of CD68 + GDF15 low macrophages (red arrowheads). The bar graphs showed the absolute and relative abundances of CD68 + GDF15 high macrophages in normal and PAH lungs. Data were expressed as mean ± SEM. * P < 0.05, unpaired t -test, n = 6.

Article Snippet: Recombinant GDF15 (#10596-GD) and recombinant interferon (IFN)-γ (#585-IF) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Immunofluorescence, Labeling

Journal: bioRxiv

Article Title: Identification of a distinct cluster of GDF15 high macrophages exhibiting anti-inflammatory activities

doi: 10.1101/2024.01.11.575315

Figure Lengend Snippet: GDF15 high macrophages could be derived by in vitro differentiation of mononuclear cells. (A and B) Immunofluorescence staining and flow cytometry results confirming that in vitro differentiation of human peripheral blood mononuclear cells (PBMNCs) with GM-CSF for 7 days yielded CD68 + macrophages. (C and D) Flow cytometry and immunofluorescence double labeling results showing that the PBMNC-derived macrophages contained a minor population of GDF15 high cells (arrowheads in D) (example from 3 independent experiments). (E and F) Fluorescence microscopy and flow cytometry data showing that GM-CSF differentiation of rat bone marrow mononuclear cells (BMMNCs) in vitro yielded macrophages (GFP expressing) of a high purity (∼90%). CD68pro-GFP rats had a GFP transgene under the control of CD68 promoter. Cells from normal rats showing no GFP fluorescence served as a negative control (left panel in E). (G and H) Flow cytometry and immunofluorescence double labeling data (from 3 independent experiments) showing that the BMMNC-derived macrophages (from CD68pro-GFP rats) contained a minor population of GDF15 high cells (arrowheads in H). The flow cytometry data in panels C and G were from cells gated for GFP + . The nuclei were counterstained with DAPI (blue).

Article Snippet: Recombinant GDF15 (#10596-GD) and recombinant interferon (IFN)-γ (#585-IF) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Derivative Assay, In Vitro, Immunofluorescence, Staining, Flow Cytometry, Labeling, Fluorescence, Microscopy, Expressing, Control, Negative Control

Journal: bioRxiv

Article Title: Identification of a distinct cluster of GDF15 high macrophages exhibiting anti-inflammatory activities

doi: 10.1101/2024.01.11.575315

Figure Lengend Snippet: Flow cytometry results showing that GDF15 high macrophages did not exhibit a typical M1 or M2 phenotype. Experiments were performed in human PBMNC-derived macrophages, using CD86, CD80 and IL-1β as the M1 markers, and CD206, CD163 and IL-4 as the M2 markers. Data were from a single test using pooled samples from 4 healthy volunteers.

Article Snippet: Recombinant GDF15 (#10596-GD) and recombinant interferon (IFN)-γ (#585-IF) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Flow Cytometry, Derivative Assay

Journal: bioRxiv

Article Title: Identification of a distinct cluster of GDF15 high macrophages exhibiting anti-inflammatory activities

doi: 10.1101/2024.01.11.575315

Figure Lengend Snippet: Molecular characterization of human PBMNC-derived GDF15 high macrophages with scRNA-seq. (A) Graphical outline of the experimental procedure. (B) UMAP plots showing the identified cell clusters (C1 to C7) based on the scRNA-seq data from total 73,768 cells combined from samples from 3 healthy volunteers, 3 PAH patients harboring mutations in BMPR2 gene, and 3 PAH patients without BMPR2 mutations. The inset showed that macrophages from healthy controls and PAH patients had virtually identical clustering profiles when analyzed separately. The putative nomenclatures for C1 to C7 were given below the graph. (C) Violin plots showing the expression patterns of identified marker genes for C1 to C7. (D) UMAP plots showing expression patterns of the top 10 genes that were overexpressed in GDF15 high macrophages (C5) as compared to GDF15 low cells. (E) UMAP plots showing expression patterns of the top 9 genes encoding secreted proteins which were overexpressed in GDF15 high macrophages as compared to GDF15 low cells. (F) Cell-cell communication network map created using CellChat showing the possible effector cells of the GDF15 high macrophage. (G and H) Predicted ligand-receptor pairs potentially involved in the signaling of reciprocal communications between GDF15 high macrophage and other cell types as listed in panel F.

Article Snippet: Recombinant GDF15 (#10596-GD) and recombinant interferon (IFN)-γ (#585-IF) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Derivative Assay, Expressing, Marker

Journal: bioRxiv

Article Title: Identification of a distinct cluster of GDF15 high macrophages exhibiting anti-inflammatory activities

doi: 10.1101/2024.01.11.575315

Figure Lengend Snippet: GDF15 high macrophages exhibited reduced inflammatory activation in vitro. (A) Expression patterns of potential substitute cell surface markers for GDF15 based on the scRNA-seq data. (B) Flow cytometry results showing that rat BMMNC-derived macrophages contained a minor fraction of TNFSF9 high cells, whose expression level was correlated with that of GDF15 (from 3 independent experiments). (C) Flow cytometry verification of the correlation between TNFSF9 and GDF15 expressions in human PBMNC-derived macrophages (from 2 independent experiments). (D) Real-time PCR results showing that GDF15 high macrophages (H) exhibited reduced expressions of TNF-α, IL-1β and IL-6 in response to LPS stimulation (1 μg/mL for 6 hr), as compared to GDF15 low cells (L). Rat BMMNC-derived macrophages were FACS purified using TNFSF9 as a substitute marker for GDF15, and primed with IFN-γ (10 ng/mL for 12 hr). (E) Boyden chamber cell migration assay showing that GDF15 high macrophages (H) exhibited reduced migratory activity as compared to GDF15 low cells (L) in the absence and presence of LPS stimulation. (F) Representative fluorescent microscopic images and quantitative data showing that GDF15 high macrophages exhibited reduced phagocytic activity in the presence of LPS stimulation as compared to GDF15 low cells. Phagocytosis was assessed by internalization of fluorochrome-labeled latex beads (orange color). The macrophages were from CD68pro-GFP rats. Data were mean ± SEM. * P < 0.05, one-way ANOVA, n = 3 in D; 6 in E; 6 in F. NS, no significance.

Article Snippet: Recombinant GDF15 (#10596-GD) and recombinant interferon (IFN)-γ (#585-IF) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Activation Assay, In Vitro, Expressing, Flow Cytometry, Derivative Assay, Real-time Polymerase Chain Reaction, Purification, Marker, Cell Migration Assay, Activity Assay, Labeling

Journal: bioRxiv

Article Title: Identification of a distinct cluster of GDF15 high macrophages exhibiting anti-inflammatory activities

doi: 10.1101/2024.01.11.575315

Figure Lengend Snippet: GDF15 high macrophages exerted anti-inflammatory effects via paracrine mechanisms. (A to C) RAW264.7 cells co-cultured with rat BMMNC-derived GDF15 high (H) or GDF15 low (L) macrophages were left untreated or stimulated with LPS for 4 hr. Results for the expression of pro-inflammatory cytokines (A, real-time PCR), cell migratory activity (B), and phagocytic activity (C) were shown. (D to F) Unsorted rat BMMNC-derived macrophages co-cultured with rat GDF15 high (H) or GDF15 low (L) macrophages were left untreated or stimulated with LPS for 4 hr. Results for the expression of pro-inflammatory cytokines (D, real-time PCR), cell migratory activity (E), and phagocytic activity (F) were shown. Data were mean ± SEM. * P < 0.05, one-way ANOVA, n = 4-5 in A; 6 in B; 3 in C; 3-4 in D; 6 in E; 6 in F. NS, no significance.

Article Snippet: Recombinant GDF15 (#10596-GD) and recombinant interferon (IFN)-γ (#585-IF) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Cell Culture, Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Activity Assay

Journal: bioRxiv

Article Title: Identification of a distinct cluster of GDF15 high macrophages exhibiting anti-inflammatory activities

doi: 10.1101/2024.01.11.575315

Figure Lengend Snippet: GDF15 might be a macrophage-derived anti-inflammatory factor. (A) Real-time PCR results showing that treatment with exogenous GDF15 (20 ng/mL) inhibited LPS- induced expression of pro-inflammatory cytokines in RAW264.7 cells. (B) Flow cytometry results showing that GDF15 treatment had no effects on phagocytosis in RAW264.7 cells without or with LPS stimulation. (C) Representative images and quantitative data of Boyden chamber assay showing that exogenous GDF15 inhibited migration of LPS-challenged RAW264.7 cells. Cells on the membrane were stained with Giemsa. (D) Effects of conditioned medium from GDF15 high macrophages (H), as compared to the medium from GDF15 low cells (L), on the expression of pro-inflammatory cytokines in RAW264.7 cells. All experiments were performed in the presence of LPS stimulation. α-GDF15, GDF15neutralizing antibody; IgG, non-specific immunoglobulin control. (E) The same experiments as those in D carried out in rat BMMNC-derived macrophages. Data were mean ± SEM. * P < 0.05, one-way ANOVA; † P < 0.05, unpaired t -test, n = 3-5 in A; 3 in B; 6 in C; 3 in D; 3 in E. NS, no significance.

Article Snippet: Recombinant GDF15 (#10596-GD) and recombinant interferon (IFN)-γ (#585-IF) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Derivative Assay, Real-time Polymerase Chain Reaction, Expressing, Flow Cytometry, Boyden Chamber Assay, Migration, Membrane, Staining, Control

Journal: bioRxiv

Article Title: Identification of a distinct cluster of GDF15 high macrophages exhibiting anti-inflammatory activities

doi: 10.1101/2024.01.11.575315

Figure Lengend Snippet: Detection of GDF15 high macrophages in various human tissues. GDF15 high macrophages (arrowheads) were identified using immunofluorescence double labeling with anti-CD68 (green color) and anti-GDF15 (red color) antibodies in (A) colon tissues from both healthy subjects and patients with ulcerative colitis, (B) kidneys (the normal peri-tumor tissue) (tested in one sample only) and (C) atherosclerotic plaques in the carotid artery (representative data from 6 independent samples showing similar results). The nuclei were counterstained with DAPI (blue). Data were mean ± SEM. NS, no significance, unpaired t -test, n = 3 for Healthy; 4 for Colitis.

Article Snippet: Recombinant GDF15 (#10596-GD) and recombinant interferon (IFN)-γ (#585-IF) were purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Immunofluorescence, Labeling