Journal: Cell Regeneration

Article Title: A single-cell hematopoietic microenvironmental atlas reveals progressive maturation of bone marrow vascular niche

doi: 10.1186/s13619-025-00265-7

Figure Lengend Snippet: Midkine was an uncharacterized microenvironmental factor. A Quantifying the type of cell–cell interaction across all stages in the atlas. B Upset plot showing HSPC as receiver between common and unique cell–cell interactions across all stages in the atlas. C Diagram depicting treatment of Mdk +/+ and Mdk −/− mouse after lethal irradiation and transplantation as well as corresponding analyzing time point. D Representative FACS contour plot and quantification about the percentage of Lin − c-Kit + sca1 + (LSK cell, Mdk +/+ = 9, Mdk −/− = 8), hematopoietic stem cell (HSC, Mdk +/+ = 9, Mdk −/− = 8) 21 days after transplantation. Error bars, mean ± SEM. p values, t- test. E Representative FACS contour plot of CD45 ( Mdk +/+ = 8, Mdk −/− = 7), CD11b ( Mdk +/+ = 8, Mdk −/− = 7) and quantification about the percentage of CD45, CD11b and Gr-1 ( Mdk +/+ = 8, Mdk −/− = 7) at 21 days after transplantation. Error bars, mean ± SEM. p values, t- test. F Representative FACS contour plot of CD45 (DMSO = 9, iMDK = 10), CD11b (DMSO = 9, iMDK = 10) and quantification about the percentage of CD45, CD11b, Gr-1(DMSO = 9, iMDK = 10) LSK (DMSO = 8, iMDK = 9), HSC (DMSO = 8, iMDK = 9) after transplantation. Error bars, mean ± SEM. p values, t- test. G Competitive repopulation assay donated by DMSO (CD45.2, n = 4) or iMDK (CD45.2, n = 4) mice bone marrow cells which were approximately 1:1.5 mixed with CD45.1 donor cells and transplanted into lethally irradiated CD45.1 host mice. Peripheral blood was analyzed at indicted time points. Error bars, mean ± SEM. p values, t- test. H FACS plot quantifying the percentage of EdU + cell percentage in LSK in both DMSO ( n = 5) and iMDK ( n = 5) mice after drug treatment. Error bars, mean ± SEM. p values, t- test. I Representative images (dish at day 8 after seeding) and quantification of CFU total cell number, CD45 number or CD11b number derived from 20,000 Lin − cell isolated from WT mice. DMSO or iMDK was added to MethoCult medium (DMSO = 6, iMDK = 4, Vehicle = 5, MDK = 5). Error bars, mean ± SEM. p values, t- test. J Control ( n = 7) or MDK ( n = 7) overexpression of HUVEC influence myeloid cell production when HUVEC were co-cultured with lineage-negative HSPC. Error bars, mean ± SEM. p values, t- test. K Representative images (dish at day 8 after seeding) and quantification of CFU total cell number (DMSO + iMDK = 4, iMDK + Mirdametinib = 4), CD45 number (DMSO + iMDK = 4, iMDK + Mirdametinib = 4) or CD11b number (DMSO + iMDK = 4, iMDK + Mirdametinib = 3) derived from 20,000 Lin − cell isolated from WT mice. DMSO + iMDK or iMDK + Mirdametinib was added to MethoCult medium. Error bars, mean ± SEM. p values, t- test

Article Snippet: Protocols and primer sequences can be provided upon request. iMDK (MCE, HY-110171; 12.8 mg/kg; in DMSO) was administrated by intraperitoneal injection (i.p.) for 4 times after irradiation.

Techniques: Irradiation, Transplantation Assay, Derivative Assay, Isolation, Control, Over Expression, Cell Culture

Journal: Theranostics

Article Title: Endothelium-specific sensing of mechanical signals drives epidermal aging through coordinating retinoid metabolism

doi: 10.7150/thno.112299

Figure Lengend Snippet: Deciphering the Mdk-Sdc4 signaling axis in cutaneous homeostasis. A. Volcano plot showing differentially expressed genes between the aMDK group and the control group in RNA-seq analysis. B. Bar graph showing Reactome enrichment analysis terms of upregulated genes in the aMDK group. C. Left panel: Venn diagram showing the overlap of upregulated genes in the aMDK group, upregulated genes in the Pyr group, and downregulated genes in aged basal cells. Right panel: Reactome enrichment analysis of intersection genes of three groups. D. Violin plot showing the expression changes of retinol metabolism genes RARA, RBP1, and RBP4 in scRNA-seq data of human facial skin, n = 3, **p < 0.01, ***p < 0.0001. E. ImageFeaturePlot showing the expression changes of RARA, RBP1, and RBP4 in spatial transcriptomics data of young and aged human skin. F. Representative immunofluorescence images showing the expression of RARA, RBP1, and RBP4 in young and aged human skin, n = 3. G. Quantitative analysis of relative fluorescence intensity of MDK and SDC4. N = 3, *p < 0.05, **p < 0.01. H. Left panel: Representative immunofluorescence images showing changes in RBP1 expression in the control and iMDK groups. Right panel: Quantitative analysis of relative fluorescence intensity of RBP1 in the control and iMDK groups. N = 3, *p < 0.05, **p < 0.01.

Article Snippet: Following dorsal hair removal with electric clippers, mice received daily subcutaneous injections (50 μL/mouse) of the following agents for 7 consecutive days: Pyrintegrin (50 μM in PBS; MedChemExpress, HY-13306); SU6656 (40 μM in PBS; MedChemExpress, HY-B0789); Recombinant RBP1 protein (100 μg/mL in PBS; MedChemExpress, HY- P71093 ); MDK signaling inhibitor (20 μM in PBS; MedChemExpress, HY-110171); Three retinoid metabolites (Rol, RP, and HPR) were sourced from TargetMol and applied at a concentration of 50 μM each; Control groups received equivalent volumes of PBS vehicle.

Techniques: Control, RNA Sequencing, Expressing, Immunofluorescence, Fluorescence

Journal: Theranostics

Article Title: Endothelium-specific sensing of mechanical signals drives epidermal aging through coordinating retinoid metabolism

doi: 10.7150/thno.112299

Figure Lengend Snippet: Mdk-Sdc4 axis orchestrates epidermal regeneration through retinoid-mediated endothelial-basal cell crosstalk. A. FeaturePlot showing the expression changes of Rbp1 in scRNA-seq data from young and aged mouse skin. B. KEGG enrichment analysis highlighting the signaling pathways specifically enriched in Rbp1-positive cells in mouse skin. C. Upper panel: Representative immunofluorescence images showing the expression of K14 and PCNA in RP-RBP1 and control groups (left), and quantitative analysis of PCNA-positive cell numbers (right). N = 3, ****p < 0.0001. Lower panel: Representative H&E staining images showing changes in the epidermis of RP-RBP1 and control groups (left), and quantitative analysis of epidermal thickness. N = 3, ***p < 0.001. D. Violin plot showing the expression levels of Rbp1, Rara, and Rbp4 in epidermal cell clusters of skin organoids derived from neonatal and aged mice. E. Schematic diagram illustrating the reprogramming of aged mouse skin organoids using all-trans retinoic acid (atRA) and all-trans retinol (atRE). F. RT-qPCR analysis comparing the expression levels of senescence-associated markers Cdkn1a and Cdkn2a, as well as epidermal growth-related genes Egfr, Mki67, and Tp63 in control, atRA, and atRE groups, n = 3, *p < 0.05, **p < 0.01, ***p < 0.001, “ns” means no significance. G. Representative immunofluorescence images showing the expression of K14, Laminin, and P63 in control, atRA, and atRE groups. H. Quantitative analysis of the number of K14-positive cell layers, Laminin signal intensity, and P63-positive cell counts, n = 3, **p < 0.01, ***p < 0.001, “ns” means no significance.

Article Snippet: Following dorsal hair removal with electric clippers, mice received daily subcutaneous injections (50 μL/mouse) of the following agents for 7 consecutive days: Pyrintegrin (50 μM in PBS; MedChemExpress, HY-13306); SU6656 (40 μM in PBS; MedChemExpress, HY-B0789); Recombinant RBP1 protein (100 μg/mL in PBS; MedChemExpress, HY- P71093 ); MDK signaling inhibitor (20 μM in PBS; MedChemExpress, HY-110171); Three retinoid metabolites (Rol, RP, and HPR) were sourced from TargetMol and applied at a concentration of 50 μM each; Control groups received equivalent volumes of PBS vehicle.

Techniques: Expressing, Protein-Protein interactions, Immunofluorescence, Control, Staining, Derivative Assay, Quantitative RT-PCR