Journal: bioRxiv

Article Title: Propranolol reduces sarcoma growth and enhances the response to anti-CTLA4 therapy by modulating the tumor microenvironment

doi: 10.1101/2021.03.11.434711

Figure Lengend Snippet: Propranolol treatment reduces MCA205 tumor angiogenesis. A and B , Quantification of angiogenic marker Vegfa (A) and Kdr (B) gene expression in whole tumor RNA from control mice and propranolol (PRO) treated mice by qRT-PCR (n=5). C , Representative CD34 (brown) IHC staining of MCA205 tumor tissue from control mice or propranolol treated mice; hematoxylin counterstain; scale bar 100µm (left panels), and 50µm (right panels). D , Dot plot showing quantification of CD34 staining by percentages of DAB+ area in 9 control mice and 11 propranolol treated mice. *p<0.05, **p<0.01 according to multiple t test with Bonferroni correction. Mean ± SEM are depicted.

Article Snippet: For CD34 immunostaining, 10 mM citrate buffer, Ph antigen retrieval, and rat anti-mouse CD34 (1:400, MEC 14.7, Novus Biologicals) were used.

Techniques: Marker, Gene Expression, Control, Quantitative RT-PCR, Immunohistochemistry, Staining

Journal: bioRxiv

Article Title: Propranolol reduces sarcoma growth and enhances the response to anti-CTLA4 therapy by modulating the tumor microenvironment

doi: 10.1101/2021.03.11.434711

Figure Lengend Snippet: Propranolol combined with anti CTLA4 increases the number of intratumoral CD8+ T cells, reduces tumor angiogenesis, and provides long lasting immune memory against MCA205 cancer cells. A and C , Representative CD8 ( A ) and CD34 ( C ) IHC staining of MCA205 tumor tissues; hematoxylin counterstain; scale bar 100µm (left panels), and 50µm (right panels). B and D , Dot plot showing quantification of CD8 ( B ), and CD34 staining ( D ). Splenocytes from cured mice were re-stimulated ex vivo with MCA205 cancer cells for 24 hours, and intracellular cytokine expressions were quantified by flow cytometry. E , Representative flow cytometry dot plots showing the expression of TNFα and IFNγ on CD4+ T cells (upper panels) or CD8+ T cells (lower panels) with memory phenotype from naïve mice (left panels) or anti-CTLA4+PRO treated mice that had shown complete tumor regression(right panels). F , Percentages of tumor reactive T cells (IFNy+ or TNFa+) with memory phenotype (CD44hi) among CD8+ or CD4+ T cells after 24-hour ex vivo re-stimulation. *p<0.05 according to multiple t test with Bonferroni correction. Mean ± SEM are depicted.

Article Snippet: For CD34 immunostaining, 10 mM citrate buffer, Ph antigen retrieval, and rat anti-mouse CD34 (1:400, MEC 14.7, Novus Biologicals) were used.

Techniques: Immunohistochemistry, Staining, Ex Vivo, Flow Cytometry, Expressing

Journal: Microscopy research and technique

Article Title: Novel Insights Into the Ultrastructural and Immunofluorescence Characteristics of Limb Skin in the Red-Eared Slider Turtle (Trachemys scripta elegans).

doi: 10.1002/jemt.24729

Figure Lengend Snippet: FIGURE 18 | Double immunofluorescence staining of PDGFRα (green) and CD34 (red) with DAPI (blue) in the skin of the red-eared slider turtle. (A, D, G) PDGFRα staining (green) with DAPI (blue). (B, E, H) CD34 staining (red) with DAPI (blue). (C, F, I) The merged images of PDGFRα (green) and CD34 (red) with DAPI (blue). (A–C) The presence of numerous telocytes (TC) in the dermis, detected by PDGFRα (green) and CD34 (red). (D–I) Higher magnification of spindle-shaped telocytes (TC) with telopods (TP). CD34 expression is slightly more intense than PDGFRα. PDGFRα (green) and CD34 (red) co-expression appear yellow in the merged images in panels (C, F, I). Scale bars: A–C = 50 μm; D–I = 10 μm.

Article Snippet: Negative controls were treated with normal serum containing isotype- specific immunoglobulins (IgG) matching the concentration and species of the primary antibodies: normal rabbit IgG (Cell Signaling Technology, USA, #2729) for ECadherin, Melan- A, Tom20, and PDGFRα; normal mouse IgG1 (Santa Cruz Biotechnology, USA, #sc- 3877) for CK14, SOX10, CD117, and α- SMA; normal rat IgG (Santa Cruz Biotechnology, USA, #sc- 2026) for CD34; and normal goat IgG (R&D Systems, USA, #AB- 108- C) for Vimentin.

Techniques: Double Immunofluorescence Staining, Staining, Expressing

Journal: Microscopy research and technique

Article Title: Novel Insights Into the Ultrastructural and Immunofluorescence Characteristics of Limb Skin in the Red-Eared Slider Turtle (Trachemys scripta elegans).

doi: 10.1002/jemt.24729

Figure Lengend Snippet: FIGURE 19 | Double immunofluorescence staining of Vimentin (green) and CD34 (red) with DAPI (blue) in the skin of the red-eared slider turtle. (A, D, G) Vimentin staining (green) with DAPI (blue). (B, E, H) CD34 staining (red) with DAPI (blue). (C, F, I) The merged images of Vimentin (green) and CD34 (red) with DAPI (blue). (A–C) The presence of telocytes (TC) in the dermis, with CD34 expression being more intense than Vimentin. (D–I) Higher magnification of spindle-shaped telocytes (TC) with telopods (TP), revealing less Vimentin expression (green) compared to the high intensity detected by CD34 (red). Some yellow staining appears due to co-expression of Vimentin (green) and CD34 (red) in panels (C, F, I). Scale bars: A–C = 25 μm; D–I = 10 μm.

Article Snippet: Negative controls were treated with normal serum containing isotype- specific immunoglobulins (IgG) matching the concentration and species of the primary antibodies: normal rabbit IgG (Cell Signaling Technology, USA, #2729) for ECadherin, Melan- A, Tom20, and PDGFRα; normal mouse IgG1 (Santa Cruz Biotechnology, USA, #sc- 3877) for CK14, SOX10, CD117, and α- SMA; normal rat IgG (Santa Cruz Biotechnology, USA, #sc- 2026) for CD34; and normal goat IgG (R&D Systems, USA, #AB- 108- C) for Vimentin.

Techniques: Double Immunofluorescence Staining, Staining, Expressing

Journal: Microscopy research and technique

Article Title: Novel Insights Into the Ultrastructural and Immunofluorescence Characteristics of Limb Skin in the Red-Eared Slider Turtle (Trachemys scripta elegans).

doi: 10.1002/jemt.24729

Figure Lengend Snippet: FIGURE 21 | Schematic representation of the red-eared slider turtle's skin, highlighting different cell types, their locations, and corresponding antibodies showing positive immunoreactivity. Keratinocytes in the epidermis are identified by E-cadherin and CK14, which show positive reactions in all keratinocytes within the epidermal layers: peri- corneous layer (PC), stratum spinosum (SS), and stratum basalis (SB). Both E-cadherin and CK14 are not expressed in the corneous material (CM). Four types of chromatophores are identified: melanocytes in the epidermis (EP) and melanophores in the dermis (D), detected by Sox10, Melan-A, and CD117. Xanthophores and iridophores, located just beneath the epidermis (EP), are detected by Sox10 and CD117. CD34, PDGFRα, and Vimentin identify telocytes in the dermis. α-SMA detects myofibroblasts in the deep dermis. The skin also reveals well-developed blood vessels in the dermis, identified by α-SMA. Numerous peripheral nerves are located in the hypodermis (HD).

Article Snippet: Negative controls were treated with normal serum containing isotype- specific immunoglobulins (IgG) matching the concentration and species of the primary antibodies: normal rabbit IgG (Cell Signaling Technology, USA, #2729) for ECadherin, Melan- A, Tom20, and PDGFRα; normal mouse IgG1 (Santa Cruz Biotechnology, USA, #sc- 3877) for CK14, SOX10, CD117, and α- SMA; normal rat IgG (Santa Cruz Biotechnology, USA, #sc- 2026) for CD34; and normal goat IgG (R&D Systems, USA, #AB- 108- C) for Vimentin.

Techniques:

Journal: Cell Transplantation

Article Title: Adipose-derived stem cells inhibit dendritic cell migration by secreting tumor necrosis factor-α-stimulated gene 6 to improve the allogeneic skin transplantation survival rate in mice

doi: 10.1177/09636897251376125

Figure Lengend Snippet: Characteristics of adipose-derived stem cells. (a) Positive expression for CD90 and CD105 and negative expression for CD19 and CD34. (b), (c) Differentiation of ADSCs into adipocytes and osteoblasts. After adipogenic differentiation, newly differentiated adipocytes had lipid droplets identified by Oil Red O staining. Osteogenic differentiation was confirmed by Alizarin Red S staining. ADSCs: adipose-derived stem cells; CD: cluster of differentiation.

Article Snippet: Passage 3 cells were stained with antibodies against CD90 (E-AB-F1283C) (Elabscience, Wuhan, China), CD105 (E-AB-F1233UE) (Elabscience, Wuhan, China), CD19 (E-AB-F1004E) (Elabscience, Wuhan, China), and CD34 (E-AB-F1284D) (Elabscience, Wuhan, China).

Techniques: Derivative Assay, Expressing, Staining

Journal: Cell Transplantation

Article Title: Adipose-derived stem cells inhibit dendritic cell migration by secreting tumor necrosis factor-α-stimulated gene 6 to improve the allogeneic skin transplantation survival rate in mice

doi: 10.1177/09636897251376125

Figure Lengend Snippet: Adipose-derived stem cells inhibited dendritic cell migration in vivo . (a) Mouse skin allotransplantation and phosphate-buffered saline (PBS) or cell infusion. (b) Expression of dendritic cells (DCs) in grafted skin. Data are expressed as fold-changes relative to levels in the control group. Statistically significant differences among groups are reported above the columns (*** P < 0.001). (c) Immunofluorescence findings. CD80 (red) and CD86 (green) are commonly used as markers for dendritic cells. The expression levels of DCs in the control group were lower than in the adipose-derived stem cell (ADSC) group. (d) Comparison of histologic changes among the two groups. Inflammatory reactions were more prominent in the control group than in the experimental group on postoperative Day 7 (×4 or ×10). CD: cluster of differentiation.

Article Snippet: Passage 3 cells were stained with antibodies against CD90 (E-AB-F1283C) (Elabscience, Wuhan, China), CD105 (E-AB-F1233UE) (Elabscience, Wuhan, China), CD19 (E-AB-F1004E) (Elabscience, Wuhan, China), and CD34 (E-AB-F1284D) (Elabscience, Wuhan, China).

Techniques: Derivative Assay, Migration, In Vivo, Saline, Expressing, Control, Immunofluorescence, Comparison

Journal: Cell Transplantation

Article Title: Adipose-derived stem cells inhibit dendritic cell migration by secreting tumor necrosis factor-α-stimulated gene 6 to improve the allogeneic skin transplantation survival rate in mice

doi: 10.1177/09636897251376125

Figure Lengend Snippet: Tumor necrosis factor-α-stimulated gene 6 played a key role in inhibiting dendritic cell maturation and migration. (a) Flow cytometry analysis of dendritic cell (DC) maturation. DCs were co-cultured with adipose-derived stem cells (ADSCs) or individual immunomodulatory factors (TSG-6, HGF, TGFβ, Galectin-1), followed by staining for CD80 and CD86. Quantification of mature DCs is shown on the right. (*** P < 0.001 vs control). (b) Transwell migration assay to assess DC motility after co-culture with ADSCs or indicated factors. Representative images (left) and quantification of migrated cells (right) (*** P < 0.001). (c) Cell Counting Kit-8 (CCK-8) assay for DC viability. CD: cluster of differentiation; TSG-6: tumor necrosis factor-α-stimulated gene 6; HGF: hepatocyte growth factor; TGFβ: transforming growth factor beta.

Article Snippet: Passage 3 cells were stained with antibodies against CD90 (E-AB-F1283C) (Elabscience, Wuhan, China), CD105 (E-AB-F1233UE) (Elabscience, Wuhan, China), CD19 (E-AB-F1004E) (Elabscience, Wuhan, China), and CD34 (E-AB-F1284D) (Elabscience, Wuhan, China).

Techniques: Migration, Flow Cytometry, Cell Culture, Derivative Assay, Staining, Control, Transwell Migration Assay, Co-Culture Assay, Cell Counting, CCK-8 Assay

Journal: Cell Transplantation

Article Title: Adipose-derived stem cells inhibit dendritic cell migration by secreting tumor necrosis factor-α-stimulated gene 6 to improve the allogeneic skin transplantation survival rate in mice

doi: 10.1177/09636897251376125

Figure Lengend Snippet: Tumor necrosis factor-α-stimulated gene 6 secreted by adipose-derived stem cells inhibited dendritic cell migration in vitro . (a, b) Tumor necrosis factor-α-stimulated gene 6 (TSG-6) effectively inhibited the migration of dendritic cells (DCs), and this inhibitory effect became more pronounced with increasing TSG-6 concentrations. (c, d) The ability of transfected adipose-derived stem cells (ADSCs) to inhibit the migration of DCs was weakened. When TSG-6 was added again, the ability of transfected ADSCs to inhibit the migration of DCs was restored (* P < 0.05; ** P < 0.01).

Article Snippet: Passage 3 cells were stained with antibodies against CD90 (E-AB-F1283C) (Elabscience, Wuhan, China), CD105 (E-AB-F1233UE) (Elabscience, Wuhan, China), CD19 (E-AB-F1004E) (Elabscience, Wuhan, China), and CD34 (E-AB-F1284D) (Elabscience, Wuhan, China).

Techniques: Derivative Assay, Migration, In Vitro, Transfection

Journal: Cell Transplantation

Article Title: Adipose-derived stem cells inhibit dendritic cell migration by secreting tumor necrosis factor-α-stimulated gene 6 to improve the allogeneic skin transplantation survival rate in mice

doi: 10.1177/09636897251376125

Figure Lengend Snippet: Transcriptomic analysis and functional enrichment of dendritic cells under different treatments. (a) Volcano plots of differentially expressed genes (DEGs) comparing ADSCs versus control (top) and TSG-6 versus control (bottom) groups. Red and blue dots indicate upregulated and downregulated genes, respectively (|log2FC| > 1, P < 0.05). (b) RT-qPCR validation of selected DEGs involved in immune modulation (ADM, GHRH, NDRG1, SELENBP1, etc.) in DCs from control, TSG-6, and ADSC groups (* P < 0.05). (c) GO enrichment analysis of DEGs in the ADSC (left) and TSG-6 (right) groups, showing the top 30 terms from biological processes (BP), molecular functions (MF), and cellular components (CC). (d) KEGG pathway enrichment of DEGs in ADSC (left) and TSG-6 (right) groups. Bubble size indicates gene count, and color represents adjusted p -values (C: control group; A: ADSC group; T: TSG-6 group). ADSC: adipose-derived stem cell; TSG-6: tumor necrosis factor-α-stimulated gene 6; RT-qPCR: real-time quantitative polymerase chain reaction; ADM: adrenomedullin; GHRH: growth hormone–releasing hormone; SELENBP1: selenium binding protein 1; NDRG1: N-myc downstream regulated gene-1; GO: gene ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: Passage 3 cells were stained with antibodies against CD90 (E-AB-F1283C) (Elabscience, Wuhan, China), CD105 (E-AB-F1233UE) (Elabscience, Wuhan, China), CD19 (E-AB-F1004E) (Elabscience, Wuhan, China), and CD34 (E-AB-F1284D) (Elabscience, Wuhan, China).

Techniques: Functional Assay, Control, Quantitative RT-PCR, Biomarker Discovery, Derivative Assay, Real-time Polymerase Chain Reaction, Binding Assay