Journal: iScience

Article Title: Single-cell RNA sequencing and spatial transcriptomics of bladder Ewing sarcoma

doi: 10.1016/j.isci.2024.110921

Figure Lengend Snippet: Bladder ES-mast is a specific type of bladder ES/PNET mast cells and promotes bladder ES/PNET progression via TNFSF12-TNFRSF12A ligand-receptor signaling pattern (A) t-SNE plots of all mast cells in 4 samples. (B) Number of mast cells in the 4 samples. (C) Re-annotated t-SNE plots of all mast cells in tumor samples. (D) Unsupervised pseudotime trajectory of 3 mast cell subtypes by RNA velocity. (E) Expression of marker genes for 3 mast cell subtypes. (F) Distribution maps of 3 mast cell subtypes in BCa_T1 and BCa_T2. (G) Pseudotime trajectory of 3 mast cell subtypes in BCa_T1 and BCa_T2. (H) Smoothed expression curves of representative candidate genes along the trajectory. (I) An overview of mast2 (bladder ES-Mast) and other cells interactions. Arrow and edge color indicate direction. Circle size is proportional to the number of cells in each cell group. Edge thickness indicates the number and the strength of interaction between populations. (J) An overview of Epi1 (bladder ES-Epi) and other 3 mast cell subtypes interactions. (K) Heatmaps of differential number and strength of intercellular interactions between Epi1 (bladder ES-Epi) and other 3 mast cell subtypes. (L) Outgoing and incoming signaling patterns of intercellular interactions between Epi1 (bladder ES-Epi) and other 3 mast cell subtypes. (M) Transcription factor analysis of 3 mast cell subtypes. (N) Ligand-receptor pair plot for Epi 1 (bladder ES-Epi) and other 3 mast cells subtypes. (O) Scatterplot of correlation between TNFRSF12A and TNFSF12 in the TCGA-BLCA database. (P) Immunofluorescence staining of TNFRSF12A in BCa_T1 and BCa_T2 (Scale bars, 100 μm). (Q) Effect of exogenous supplementation with TNFSF12 or Enavatuzumab on the migratory capacity of RD-ES cells. (Scale bars, 100 μm, n = 3, Data were analyzed with use of Student’s t test, and represented as mean ± SEM, ∗∗∗ p < 0.001). (R) Localization of epithelial cells and mast cells, as well as TNFRSF12A and TNFSF12 at the ST level.

Article Snippet: Enavatuzumab , MCE , HY-P99361.

Techniques: Expressing, Marker, Immunofluorescence, Staining

Journal: iScience

Article Title: Single-cell RNA sequencing and spatial transcriptomics of bladder Ewing sarcoma

doi: 10.1016/j.isci.2024.110921

Figure Lengend Snippet:

Article Snippet: Enavatuzumab , MCE , HY-P99361.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Software

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Expression of TWEAK and Its Receptor Fn14 in the Multiple Sclerosis Brain: Implications for Inflammatory Tissue Injury

doi: 10.1097/nen.0b013e31818dab90

Figure Lengend Snippet: FIGURE 5. Detection of Fn-14 in the brain of control and MS cases. (A) Immunostaining with anti-Fn14 antibody shows that the normal white matter is largely negative for this molecule; the inset highlights the occasional finding of isolated cells with an astrocytic morphology that are strongly immunoreactive for Fn14. (BYG) Immunostaining performed in highly inflammatory brains from secondary progressive multiple sclerosis (SPMS) cases with ectopic follicles. (B) Immunostaining reveals the presence of several Fn14+ reactive astrocytes at the edge of a white matter lesion; the inset shows one of the rare Fn14+ cells with a microglia-like morphology identified in the same lesion. (C) Some astrocytes are Fn14+ in a subpial lesion close to an ectopic follicle. (D, E) In 2 different SPMS cases with marked meningeal inflammation, Fn14 immunoreactivity is present on some neuronal cell bodies (D) and processes (E). (F, G) Double immunofluorescence staining with anti-Fn14 monoclonal antibody (green) and anti-NF 200 (red) demonstrates Fn14 immunoreactivity on the membrane of neurofilament+ axons. Original magnification: (A, B, D) 500; ([C] and inset in [A]) 1,000. Scale bars = (E) 50 Km; (F) 20 Km.

Article Snippet: Air-dried acetone-fixed 10-Km-thick cryosections were immunostained with the following antibodies (Abs): antiTWEAK monoclonal Ab (mAb) (clone P2D10; Biogen Idec, Cambridge, MA), and anti-Fn14 mAb (clone ITEM4; Acris Antibodies, Herford, Germany), anti-Iba-1 (ionized calcium binding adapter molecule-1) rabbit polyclonal Ab (a kind gift of Dr Y. Imai, Tokyo, Japan) for macrophages/microglia; anti-CD20 for B cells (Immunotech, Marseille, France); antiimmunoglobulin A, M, G polyclonal Ab (Dako, Carpinteria, CA) for plasma cells, anti-HLA-DR mAb (CR3/43, Dako), anti-MOG mAb (Serotec, Oxford, United Kingdom) for myelin staining, anti-glial fibrillary acidic protein rabbit polyclonal antibody (Dako) and mAb (Biogenex, San Ramon, CA) for astrocytes, anti-cleaved caspase-3 rabbit polyclonal Ab (Cell Signaling Technology, Boston, MA) to stain apoptotic cells; anti-CD31 mAb for endothelial cells (Dako), anti-laminin rabbit polyclonal Ab for the basal membrane (Dako), and anti-neurofilament (200 kd) rabbit polyclonal antibody for neurons (DBA, Segrate, Milan, Italy).

Techniques: Control, Immunostaining, Isolation, Staining, Membrane

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: Expression of TWEAK and Its Receptor Fn14 in the Multiple Sclerosis Brain: Implications for Inflammatory Tissue Injury

doi: 10.1097/nen.0b013e31818dab90

Figure Lengend Snippet: FIGURE 6. TWEAK and Fn14 gene expression in control and MS cortex. TWEAK (A) and Fn14 (B) messenger RNA (mRNA) was quantitated by real-time reverse transcription-polymerase chain reaction in cortical samples dissected from brains of control (n = 9) and multiple sclerosis (MS) cases (n = 16). Data are expressed as mRNA levels normalized to glyceraldehyde-3- phosphate dehydrogenase relative to a sample of pooled control cases used for calibration. Dots represent values for each control or MS case analyzed; bars represent median values. Values of p were calculated with Mann-Whitney U test.

Article Snippet: Air-dried acetone-fixed 10-Km-thick cryosections were immunostained with the following antibodies (Abs): antiTWEAK monoclonal Ab (mAb) (clone P2D10; Biogen Idec, Cambridge, MA), and anti-Fn14 mAb (clone ITEM4; Acris Antibodies, Herford, Germany), anti-Iba-1 (ionized calcium binding adapter molecule-1) rabbit polyclonal Ab (a kind gift of Dr Y. Imai, Tokyo, Japan) for macrophages/microglia; anti-CD20 for B cells (Immunotech, Marseille, France); antiimmunoglobulin A, M, G polyclonal Ab (Dako, Carpinteria, CA) for plasma cells, anti-HLA-DR mAb (CR3/43, Dako), anti-MOG mAb (Serotec, Oxford, United Kingdom) for myelin staining, anti-glial fibrillary acidic protein rabbit polyclonal antibody (Dako) and mAb (Biogenex, San Ramon, CA) for astrocytes, anti-cleaved caspase-3 rabbit polyclonal Ab (Cell Signaling Technology, Boston, MA) to stain apoptotic cells; anti-CD31 mAb for endothelial cells (Dako), anti-laminin rabbit polyclonal Ab for the basal membrane (Dako), and anti-neurofilament (200 kd) rabbit polyclonal antibody for neurons (DBA, Segrate, Milan, Italy).

Techniques: Gene Expression, Control, Reverse Transcription, Polymerase Chain Reaction, MANN-WHITNEY

Journal: Stem Cell Reviews and Reports

Article Title: Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

doi: 10.1007/s12015-024-10695-6

Figure Lengend Snippet: Sequences of primers for each molecular gene amplification

Article Snippet: Recombinant human FN1 (Solarbio, P00167) was reconstituted in PBS containing 0.1% BSA to a working concentration of 50 ng/mL.

Techniques:

Journal: Stem Cell Reviews and Reports

Article Title: Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

doi: 10.1007/s12015-024-10695-6

Figure Lengend Snippet: Differentially expression of FN1/ITGA5 in dental pulp tissue. A : GO analysis of gene expression in DPSCs from mature and young permanent dental pulp tissues. B : Immunofluorescence staining of CD31 (red) and ITGA5 (green) in mature and young permanent dental pulp tissues. C : Immunofluorescence staining of CD31 (red) and FN1 (green) in mature and young permanent dental pulp tissues. D–G: Western blot, RT-PCR, and ELISA were used to detect FN1 expression in A-DPSCs and Y-DPSCs (* P < 0.05, ** P < 0.01, *** P < 0.001)

Article Snippet: Recombinant human FN1 (Solarbio, P00167) was reconstituted in PBS containing 0.1% BSA to a working concentration of 50 ng/mL.

Techniques: Expressing, Gene Expression, Immunofluorescence, Staining, Western Blot, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal: Stem Cell Reviews and Reports

Article Title: Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

doi: 10.1007/s12015-024-10695-6

Figure Lengend Snippet: Regulatory effects of FN1 in DPSCs on the biological functions of co-cultured HUVECs. A – D : After silencing FN1 in Y-DPSCs, western blot, RT-PCR, and ELISA were detected to validate FN1 expression; E – F : Cell migration of HUVECs co-cultured with DPSCs in a scratch assay. G – I : The tube formation assay was performed by counting branch points and capillary length. J : Proliferative ability was determined using CCK-8 assay (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: Recombinant human FN1 (Solarbio, P00167) was reconstituted in PBS containing 0.1% BSA to a working concentration of 50 ng/mL.

Techniques: Cell Culture, Western Blot, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing, Migration, Wound Healing Assay, Tube Formation Assay, CCK-8 Assay

Journal: Stem Cell Reviews and Reports

Article Title: Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

doi: 10.1007/s12015-024-10695-6

Figure Lengend Snippet: FN1 regulates the tube formation function of HUVECs via ITGA5. A / B : Cell migration of HUVECs treatment with FN1 or siR-ITGA5 in scratch assay. C – E : The tube formation assay of HUVECs treatment with FN1 or siR-ITGA5 was performed by counting branch points and capillary length. F : Proliferative ability was determined using CCK-8 assay (* P < 0.05, ** P < 0.01, *** P < 0.001)

Article Snippet: Recombinant human FN1 (Solarbio, P00167) was reconstituted in PBS containing 0.1% BSA to a working concentration of 50 ng/mL.

Techniques: Migration, Wound Healing Assay, Tube Formation Assay, CCK-8 Assay

Journal: Stem Cell Reviews and Reports

Article Title: Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

doi: 10.1007/s12015-024-10695-6

Figure Lengend Snippet: Silencing FN1 or ITGA5 in dental pulp tissue exerts inhibitory effects on dental pulp vascular development. A : Inhibitory modulation of FN1 and ITGA5 functionality in developing dental pulp tissue was achieved through periodontal ligament injections of siRNA targeting FN1 or ITGA5, respectively, in mice. B : Immunofluorescence staining of CD31 (red) and FN1 or ITGA5 (green) in mature and young permanent dental pulp tissues. C : Numbers of vessels in dental pulp tissues of each group (* P < 0.05, ** P < 0.01, *** P < 0.001)

Article Snippet: Recombinant human FN1 (Solarbio, P00167) was reconstituted in PBS containing 0.1% BSA to a working concentration of 50 ng/mL.

Techniques: Immunofluorescence, Staining

Journal: Stem Cell Reviews and Reports

Article Title: Enhancing Vasculogenesis in Dental Pulp Development: DPSCs-ECs Communication via FN1-ITGA5 Signaling

doi: 10.1007/s12015-024-10695-6

Figure Lengend Snippet: FN1/ITGA5 regulates the tube formation function of HUVECs via the PI3K/AKT signaling pathway. A / B : Evaluation of PI3K/AKT signaling pathway activation status in HUVECs treated with FN1 or siRNA using Western blot. C / D : Evaluation of PI3K/AKT signaling pathway activation status in HUVECs treated with FN1 or LY294002 using Western blot. E / F : Cell migration of HUVECs treatment with FN1 or LY294002 in scratch assay. G – I : The tube formation assay of HUVECs treatment with FN1 or LY294002 was performed by counting branch points and capillary length (* P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001)

Article Snippet: Recombinant human FN1 (Solarbio, P00167) was reconstituted in PBS containing 0.1% BSA to a working concentration of 50 ng/mL.

Techniques: Activation Assay, Western Blot, Migration, Wound Healing Assay, Tube Formation Assay