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