recombination human resistin  (PeproTech)

Journal: Nature Communications

Article Title: The REDD1–NF-κB–miRNAs–eNOS/SIRT1 axis mediates obesity-induced endothelial cell senescence and hypertension

doi: 10.1038/s41467-026-70601-1

Figure Lengend Snippet: a – c Aortic vessels were isolated from male mice fed either a chow diet (lean) or an HFD (DIO) for 16 weeks. Vessels were either intact (E+) or de-endothelialized (E−). REDD1 expression was assessed by Western blotting ( a , b ) and confocal microscopy ( c ). Representative images from four mice per group with similar results. Scale bar = 50 µm. d , e REDD1 mRNA ( d , n = 4 independent experiments) and REDD1 protein levels ( e , n = 3 independent experiments) were measured after treatment with BSA-conjugated palmitic acid (PA, 300 µM), cholesterol (Chol, 100 µM, dissolved in DMSO), leptin (10 µg/ml), resistin (Retn, 100 ng/ml), oxLDL (50 µg/ml), or high glucose (HG, 25 mM) using qRT-PCR and Western blotting. f – h HAECs were transfected with 80 nM control siRNA (siC) or REDD1 siRNA (siREDD1) and then treated with PA ( f ), oxLDL ( g ), or high glucose (HG, h ) for 48 h. SA-β-gal + cells were detected using a SA-β-gal staining kit. Scale bar = 100 µm. The percentage of SA-β-gal + cells was calculated as the ratio of blue-stained to total cells ( n = 4 independent experiments). Western blotting for REDD1, p53, and p21 was performed in three independent experiments with similar results. Data are presented as mean ± s.e.m. Statistical significance was determined using one-way ANOVA with Holm–Sidak’s multiple comparisons test ( d , f – h ).

Article Snippet: Cells were then treated with or without oxLDL (50 μg/ml), prepared by incubating native LDL (1.019 mg protein) with 10 μM CuSO4 for 4 h , cholesterol (100 μM, #C8667; Sigma-Aldrich, St. Louis, MO, USA), glucose (25 mM, 25 mM L -glucose used for osmotic control), recombinant human resistin (100 ng/ml, #1359-RN; R&D systems, Minneapolis, MN, USA), recombinant human leptin (10 μg/ml, #398-LP; R&D Systems), recombinant human TNF-α (10 ng/ml, #10291-TA; R&D Systems), recombinant human IL-6 (40 ng/ml, #206-IL; R&D Systems), palmitate-BSA (300 μM), or H 2 O 2 (50–150 μM) for 48 h. Palmitic acid-BSA solution (5 mM) was prepared by mixing 10 μL of palmitate solution (500 mM in ethanol, #P9767; Sigma-Aldrich) with 1 mL of BSA solution (10%, w/v in serum-free M199, #A8806; Sigma-Aldrich).

Techniques: Isolation, Expressing, Western Blot, Confocal Microscopy, Quantitative RT-PCR, Transfection, Control, Staining

Journal: Nature Communications

Article Title: The REDD1–NF-κB–miRNAs–eNOS/SIRT1 axis mediates obesity-induced endothelial cell senescence and hypertension

doi: 10.1038/s41467-026-70601-1

Figure Lengend Snippet: a – c Representative H&E-stained images of renal tissues from lean and DIO mice: WT and Redd1 −/− ( a , n = 8 mice/group), Redd1 fl/fl and Redd1 ΔEC ( b , n = 7 mice/group), or WT and miR-214-3p −/− ( c , n = 6 mice/group). Glomerular size was quantified in four randomly selected fields per section using ImageJ. Scale bar = 50 µm. d , e Fibrotic area in renal tissues (Masson′s trichrome staining; ( d )) and serum creatinine levels ( e ) were quantified using ImageJ and ELISA, respectively ( n = 8, 7, and 6 mice for Redd1 −/− , Redd1 ΔEC , miR-214-3p −/− groups, respectively; same mice as in ( a – c )). f Representative Western blots using target-specific antibodies from four mice per group with similar results. Target proteins were analyzed independently three times, with tubulin probed on the same membrane as a loading control. g Graphical representation illustrating the role of REDD1 in obesity-induced vascular senescence and hypertension based on our current and previous findings . FFA free fatty acid, IR insulin resistance, TG triglyceride, VLDL very-low-density lipoprotein, LPL lipoprotein lipase, CE cholesteryl ester, ROS reactive oxygen species, Glc glucose, Retn resistin, GLG glycogen. Data are presented as mean ± s.e.m. Statistical significance was determined using two-way ANOVA with Holm–Sidak’s multiple comparisons test ( a – e ).

Article Snippet: Cells were then treated with or without oxLDL (50 μg/ml), prepared by incubating native LDL (1.019 mg protein) with 10 μM CuSO4 for 4 h , cholesterol (100 μM, #C8667; Sigma-Aldrich, St. Louis, MO, USA), glucose (25 mM, 25 mM L -glucose used for osmotic control), recombinant human resistin (100 ng/ml, #1359-RN; R&D systems, Minneapolis, MN, USA), recombinant human leptin (10 μg/ml, #398-LP; R&D Systems), recombinant human TNF-α (10 ng/ml, #10291-TA; R&D Systems), recombinant human IL-6 (40 ng/ml, #206-IL; R&D Systems), palmitate-BSA (300 μM), or H 2 O 2 (50–150 μM) for 48 h. Palmitic acid-BSA solution (5 mM) was prepared by mixing 10 μL of palmitate solution (500 mM in ethanol, #P9767; Sigma-Aldrich) with 1 mL of BSA solution (10%, w/v in serum-free M199, #A8806; Sigma-Aldrich).

Techniques: Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Membrane, Control

Journal: bioRxiv

Article Title: Bone Marrow Adipokine Mediates Hematopoietic Regeneration and Stem Cell Fitness

doi: 10.1101/2025.08.28.672647

Figure Lengend Snippet: (a) Heat map showing the differentially regulated pathways in SLAMs sorted from WT and Adipo-RXRα/β Δ/Δ mice. (b) Pathway analysis with downregulated DEGs (P≤0.05, FC ≤−1.5). (c) Pathway enrichment analysis of different clusters with differentially regulated genes. (d) UMAP plot with different population clusters in WT and Adipo-RXRαβ Δ/Δ derived LSK cells. (e) Number of differentially regulated genes in different population clusters. (f) Volcano plot of differentially regulated genes in three different clusters of HSCP, MPP and LMPP. (g) Pathway enrichment analysis of different clusters with differentially regulated genes from scRNASeq analysis. (h) Heatmap of different adipokine levels in bone marrow extracellular fluid of WT and Adipo-RXRαβ Δ/Δ mice. (i-j) Quantification of Resistin concentration in BM extracellular fluid and serum in WT and Adipo-RXRαβ Δ/Δ mice. Data are presented as mean ± SD. Unpaired t-test was performed for statistical analysis. *p<0.05; **p<0.01; ***p <0.001.

Article Snippet: Recombinant Resistin was purchased form R&D Systems (Minneapolis, MN).

Techniques: Derivative Assay, Concentration Assay

Journal: bioRxiv

Article Title: Bone Marrow Adipokine Mediates Hematopoietic Regeneration and Stem Cell Fitness

doi: 10.1101/2025.08.28.672647

Figure Lengend Snippet: Related to . (a) Confocal images of WT proximal BM TdTomato expressing adipocytes and preadipocytes in the proximal femur and (b) tibia. (c, d) Schematic representation of TdTomato+ (AdipoQ+) cell isolation with frequency. (e) Resistin mRNA quantification in the TdTomato+ (AdipoQ+) bone marrow stromal cells from WT mice and WT bone marrow adipocytes. (f) Schematic representation of MSC-derived adipocyte production ex vivo, RXRαβ Δ/Δ adipocyte production, conditioned media, and cell mRNA isolation for Resistin quantification. (g) Resistin mRNA quantification. (h) Resistin concentration in the conditioned media. (i) Schematic representation of Resistin neutralization experiment. Quantification of (j) blood chimera, (k) bone marrow cellularity, (l) donor chimerism in bone marrow, (m) B, myeloid and T cell numbers in bone marrow in primary recipients. (n, o) Donor chimerism, B, myeloid, and T cell numbers in the blood of secondary recipients. Quantification of the (p) bone marrow cellularity, (q) donor chimerism in bone marrow, (r) B, myeloid, and T cell numbers in bone marrow in secondary recipients. (s-x) Quantification of different HSC/P populations. Data are presented as mean ± SD. Unpaired t-test and two-way ANOVA were performed for statistical analysis. *p<0.05; **p<0.01; ***p <0.001.

Article Snippet: Recombinant Resistin was purchased form R&D Systems (Minneapolis, MN).

Techniques: Expressing, Cell Isolation, Derivative Assay, Ex Vivo, Isolation, Concentration Assay, Neutralization

Journal: bioRxiv

Article Title: Bone Marrow Adipokine Mediates Hematopoietic Regeneration and Stem Cell Fitness

doi: 10.1101/2025.08.28.672647

Figure Lengend Snippet: (a) Schematic representation of ex vivo Resistin treatment to WT LSK cells and serial competitive repopulation assay. (b-e) Quantification of blood chimera, myeloid, B and T cell repopulation after serial competitive transplant. (f-k) Quantification of bone marrow chimera; B, myeloid and T cell number after serial competitive transplant. (l) Quantification of CFU-C after ex vivo Resistin treatment to WT LSK. (m) Flowcytometric plot for Pyronin Y-DAPI staining. (n) Quantification of cell frequency in different cell cycle stages. (o) Heat map showing the differentially regulated pathways in SLAMs sorted from WT and Adipo-RXRα/β Δ/Δ mice. (p) DEGs related to NF-kB signaling. (q) Pathway analysis with upregulated DEGs (P≤0.05, FC ≤−1.2). Data are presented as mean ± SD. Unpaired t-test, two-way ANOVA and chi square test were performed for statistical analysis. *p<0.05; **p<0.01; ***p <0.001.

Article Snippet: Recombinant Resistin was purchased form R&D Systems (Minneapolis, MN).

Techniques: Ex Vivo, Staining

Journal: bioRxiv

Article Title: Bone Marrow Adipokine Mediates Hematopoietic Regeneration and Stem Cell Fitness

doi: 10.1101/2025.08.28.672647

Figure Lengend Snippet: Related to . (a) Bone marrow cellularity of primary and secondary recipients. (b) WBC (Leucocyte) count and (c) donor chimerism of primary and secondary recipients. (d-i) Donor chimerism, bone marrow cellularity, B, myeloid and T cell number of primary and secondary recipients. (j) Confocal microscopic images of HSCs with NF-kB staining. (k) Quantification of the NF-kB nuclear localization after Resistin treatment. Scale bar=5 μm. Data are presented as mean ± SD. Unpaired t-test and two-way ANOVA were performed for statistical analysis. *p<0.05; **p<0.01; ***p <0.001.

Article Snippet: Recombinant Resistin was purchased form R&D Systems (Minneapolis, MN).

Techniques: Staining

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: Induction of IL‐20 expression in NP cells by resistin stimulation. RNA samples were isolated at the indicated time points or doses and subjected to real‐time polymerase chain reaction analysis. Data are presented as fold changes in fluorescent density from control NP cells (CL) normalized to 18S rRNA level (A, B). The IL‐20 protein in conditioned media was detected by enzyme‐linked immunosorbent assay (C, D). NP cells were kept as controls (CL) or stimulated with 50 ng/mL resistin for the times indicated (C), or the cells were stimulated with resistin at various doses for 4 h (D). Data are shown as mean ± standard error of the mean (SEM). * p < 0.05 versus control NP cells (CL).

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Expressing, Isolation, Real-time Polymerase Chain Reaction, Control, Enzyme-linked Immunosorbent Assay

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: p38 MAPK and Akt pathways are required for resistin‐induced expression of IL‐20. (A, B) NP cells were kept as control (CL) or stimulated with 50 ng/mL resistin for 2 h (A) and 4 h (B). Before being kept as CL or stimulated with resistin, NP cells were pretreated with PD98059 (PD), SP600125 (SP), SB203580 (SB), or LY294002 (LY) individually for 1 h. (A) RNA was isolated and subjected to real‐time polymerase chain reaction analysis. Data are presented as fold changes in fluorescent density from control NP cells (CL) normalized to 18S rRNA level. (B) The IL‐20 protein secretion in conditioned media was determined by enzyme‐linked immunosorbent assay. The results are shown as mean ± standard error of the mean. * p < 0.05 versus CL. # p < 0.05 versus vehicle control (dimethyl sulfoxide) with resistin stimulation. (C) NP cells were employed as CL or stimulated with resistin for the durations indicated, and p38 and Akt phosphorylations were determined by Western blotting.

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Expressing, Control, Isolation, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: Induction of NF‐kB‐p65 activity by resistin stimulation in NP cells. (A) IL‐20 mRNA expression levels were determined in NP cells pretreated with vehicle (DMSO) or SN50, or transfected with control siRNA (si‐CL) or si‐p65, and then stimulated with 50 ng/mL resistin for 2 h. RNA samples were isolated and subjected to real‐time polymerase chain reaction analysis. Data are presented as fold changes in fluorescent density from control NP cells (CL) normalized to 18S rRNA level. (B, C) The NF‐kB p65 activation was determined by a transcription factor (TF)‐enzyme‐linked immunosorbent assay. (B) NP cells were employed as control (CL) or stimulated with resistin for the durations indicated. (C) NP cells were pretreated individually with PD98059 (PD), SP600125 (SP), SB203580 (SB), or LY294002 (LY) for 1 h before being used as controls (CL) or stimulated with 50 ng/mL resistin for 1 h. NF‐κB p65 activity was then analyzed. All bar graphs represent folds of CL NP cells, mean ± standard error of the mean. * p < 0.05 versus CL. # p < 0.05 versus DMSO or si‐CL under resistin stimulation.

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Activity Assay, Expressing, Transfection, Control, Isolation, Real-time Polymerase Chain Reaction, Activation Assay, Enzyme-linked Immunosorbent Assay

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: Blockade of TLR4 activity inhibited resistin‐induced IL‐20 expression. NP cells were kept as controls (CL), or pretreated with isotype‐matched IgG (Ab‐IgG) and specific TLR4 neutralizing antibody (Ab‐TLR4), or transfected with the control siRNA (si‐CL) and si‐TLR4, and subsequently stimulated with resistin for 2 h (A) and 1 h (B). (A) IL‐20 mRNA levels were determined through real‐time PCR in NP cells and normalized to 18S rRNA. (B) The activation of NF‐kB‐p65 in NP cells after resistin stimulation was analyzed using transcription factor (TF) ELISA. All bar graphs represent folds of control NP cells (CL), mean ± standard error of the mean. * p < 0.05 versus CL NP cells. # p < 0.05 versus IgG‐pretreated or si‐CL‐transfected NP cells under resistin stimulation.

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Activity Assay, Expressing, Transfection, Control, Real-time Polymerase Chain Reaction, Activation Assay, Enzyme-linked Immunosorbent Assay

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: Pre‐exposure of NP cells to 5% cyclic stretch with 0.1 Hz for 30 min inhibited resistin‐induced IL‐20 expression. Static NP cells were stimulated with resistin without prestretching (static). NP cells were kept as controls (CL) or pre‐exposed to cyclic stretch at 5% with 0.1 Hz for the indicated durations followed by resistin stimulation. (A) The mRNA levels of IL‐20 in NP cells were determined through real‐time polymerase chain reaction and normalized to 18S rRNA. * p < 0.05 versus CL NP cells. ** p < 0.05 versus static NP cells with resistin stimulation. # p < 0.05 versus resistin‐treated NP cells with cyclic stretch at 5% with 0.1 Hz for 10' and 1 h. (B) The phosphorylation of p38 MAPK and Akt was determined by Western blotting. (C) NF‐kB‐p65 activation in NP cells after 1 h resistin stimulation was analyzed by TF‐ELISA. All bar graphs represent folds of control NP cells (CL), mean ± standard error of the mean. * p < 0.05 versus CL NP cells. # p < 0.05 versus static NP cells with resistin stimulation.

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Phospho-proteomics, Western Blot, Activation Assay, Enzyme-linked Immunosorbent Assay, Control

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: Upregulation of NRF2 inhibited resistin‐induced IL‐20 expression in NP cells. (A) NP cells were used as static control (static) or exposed to 5% with 0.1 Hz cyclic stretch for 30 min or 2 h. The expression of NRF2 in the nucleus was determined by Western blotting. (B–D) NP cells were used as static control (static), or pre‐exposed to 5% with 0.1 Hz for 30 min, and then treated with resistin (50 ng/mL) for 2 h (B), 30 min (C), and 1 h (D). Prior to cyclic stretch exposure, NP cells were transfected with the control siRNA (si‐CL) or si‐NRF2. (B) The levels of IL‐20 mRNA in NP cells were determined through real‐time polymerase chain reaction and normalized to 18S rRNA. (C) The phosphorylation of p38 MAPK, and Akt was determined by Western blotting. (D) NF‐kB‐p65 activation in NP cells was analyzed by TF‐ELISA. All bar graphs represent the percentage of static NP cells (static), mean ± standard error of the mean. * p < 0.05 versus static NP cells.

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Expressing, Control, Western Blot, Transfection, Real-time Polymerase Chain Reaction, Phospho-proteomics, Activation Assay, Enzyme-linked Immunosorbent Assay

Journal: JOR Spine

Article Title: Low‐Frequency Cyclic Stretch Upregulates the Expression of Nuclear Factor Erythroid 2‐Related Factor 2 in Human Nucleus Pulposus Cells to Inhibit the Resistin‐Induced Interleukin‐20 Expression

doi: 10.1002/jsp2.70040

Figure Lengend Snippet: Schematic representation of the signaling pathways regulating 5% with 0.1 Hz cyclic stretch‐induced NRF2 expression and consequent inhibition of resistin effect in human NP cells.

Article Snippet: Recombination human resistin was purchased from PeproTech (Rocky Hill, NJ, USA) and dissolved in deionized water to prepare a stock solution of 50 μg/mL.

Techniques: Protein-Protein interactions, Expressing, Inhibition

Journal: Journal of Translational Medicine

Article Title: Multi-omics analysis reveals a feedback loop amplifying immune responses in acute graft-versus-host disease due to imbalanced gut microbiota and bile acid metabolism

doi: 10.1186/s12967-024-05577-x

Figure Lengend Snippet: Granulocyte-like cells derived from patients with non-aGVHD exhibited elevated levels of RETN expression, suggesting a potential role for resistin in preventing aGVHD. A Heatmap display of the communication probability of each cluster within the combined dataset of PBMCs from patients undergoing allo-HSCT. B RETN and its corresponding receptor CAP1 were expresed in granulocyte-like cells across each sample. C Representative immunofluorescence staining of CD45 and resistin in the small intestine of control or aGVHD mouse model groups. Scale bar, 100 µm. D IL-1β concentrations were measured using ELISA assay in cell culture supernatants (n = 3) following various treatments involving LPS and resistin. E Venn diagrams illustrating the shared conserved downregulated genes among distinct lineages (group 1: neutrophils vs. macrophages vs. monocytes; group 2: cDC1 vs. cDC2 vs. migratory DC; group 3: plasmacytoid dendritic cell (pDC) vs. B cells; group 4: CD4 + T cells vs. CD8 + T cells vs. Treg; group 5: innate lymphoid cell (ILC) vs. γδ T cells vs. NK cells). F Summary of the resistin-driven immune regulation in the presence of LPS or other special conditions

Article Snippet: Following PMA stimulation, the culture medium was replaced with RPMI1640 supplemented with 10% FBS, 1 × P/S, and LPS (1, 10, or 100 ng/mL) (Cat# L8880, Salarbio) or recombinant human resistin (1, 10, and 100 ng/mL) (Cat# CJ48, novoprotein) for 24 h. The resulting cell-free supernatants were harvested and centrifuged at 500× g for 10 min.

Techniques: Derivative Assay, Expressing, Immunofluorescence, Staining, Control, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Journal of Translational Medicine

Article Title: Multi-omics analysis reveals a feedback loop amplifying immune responses in acute graft-versus-host disease due to imbalanced gut microbiota and bile acid metabolism

doi: 10.1186/s12967-024-05577-x

Figure Lengend Snippet: A simplified schematic diagram illustrating the dysregulated gut microbiota and bile acid metabolism positive feedback loop, which exacerbates immune responses and exacerbates aGVHD. Improper treatment of patients undergoing allo-HSCT alters the gut microbiota and disrupts the imbalanced bile acid metabolism. Dysregulated bile acid metabolism results in reduced production of IL1RN, defensins, and resistin. Deficiency of defensins and resistin fails to inhibit harmful microorganisms, leading to increased production of proinflammatory cytokines and proteins, such as IL-17A, IL-1β, TNFα, IL-6, IL-12, and members of the S100A family. Additionally, it was determined that resistin can inhibit aGVHD by downregulating the expression of IL1B, DUSP1, and members of the AP-1 family

Article Snippet: Following PMA stimulation, the culture medium was replaced with RPMI1640 supplemented with 10% FBS, 1 × P/S, and LPS (1, 10, or 100 ng/mL) (Cat# L8880, Salarbio) or recombinant human resistin (1, 10, and 100 ng/mL) (Cat# CJ48, novoprotein) for 24 h. The resulting cell-free supernatants were harvested and centrifuged at 500× g for 10 min.

Techniques: Expressing