Journal: Oncology Reports

Article Title: Exosomal PD‑L1 promotes the formation of an immunosuppressive microenvironment in gastric diffuse large B‑cell lymphoma

doi: 10.3892/or.2023.8525

Figure Lengend Snippet: Protein expression levels of protein markers of B cells and the immune microenvironment in tissue specimens of gastric diffuse large B-cell lymphoma. (A and B) Immunohistochemistry images of protein expression levels of the proliferative B-cell markers CD20, CD79a, CD5, CD10, Ki67 and PD-L1 as well as the immune microenvironmental markers CD4, CD8 and PD-1 in two representative tissue specimens of GDLBCL. Scale bar=50 µm. PD-L1, programmed death-ligand 1.

Article Snippet: The antibodies used for IHC staining were as follows: Rabbit anti-PD-1 (1:200, clone D4W2J, cat. no. 86163S, Cell Signaling Technologies, Inc.), rabbit anti-PD-L1 (1:500, 17952-1-AP, Proteintech Group, Inc.), mouse anti-CD8 (1:5,000, clone 1G2B10, cat. no. 66868-1-Ig, Proteintech Group, Inc.), rabbit anti-CD5 (1:800, clone E8X3S, cat. no. 39300S, Cell Signaling Technologies, Inc.), rabbit anti-CD10 (1:500, clone E5P7S, cat. no. 65534S, Cell Signaling Technologies, Inc.), rabbit anti-CD20 (1:200, clone E7B7T, cat. no. 48750S, Cell Signaling Technologies, Inc.), rabbit anti-CD79a (1:250, clone D1X5C, cat. no. 13333S, Cell Signaling Technologies, Inc.), mouse anti-Ki67 (1:2,000, clone 8D5, cat. no. 9449S, Cell Signaling Technologies, Inc.), goat anti-mouse IgG (HRP-linked, 1:2,000, cat. no. AP124P, MilliporeSigma), and goat anti-rabbit IgG (HRP-linked, 1:2,000, cat. no. AP132P, MilliporeSigma).

Techniques: Expressing, Immunohistochemistry

Journal: Oncology Reports

Article Title: Exosomal PD‑L1 promotes the formation of an immunosuppressive microenvironment in gastric diffuse large B‑cell lymphoma

doi: 10.3892/or.2023.8525

Figure Lengend Snippet: The relationship between the protein level of PD-L1 in plasma exosomes and the immune microenvironment of gastric diffuse large B-cell lymphoma.

Article Snippet: The antibodies used for IHC staining were as follows: Rabbit anti-PD-1 (1:200, clone D4W2J, cat. no. 86163S, Cell Signaling Technologies, Inc.), rabbit anti-PD-L1 (1:500, 17952-1-AP, Proteintech Group, Inc.), mouse anti-CD8 (1:5,000, clone 1G2B10, cat. no. 66868-1-Ig, Proteintech Group, Inc.), rabbit anti-CD5 (1:800, clone E8X3S, cat. no. 39300S, Cell Signaling Technologies, Inc.), rabbit anti-CD10 (1:500, clone E5P7S, cat. no. 65534S, Cell Signaling Technologies, Inc.), rabbit anti-CD20 (1:200, clone E7B7T, cat. no. 48750S, Cell Signaling Technologies, Inc.), rabbit anti-CD79a (1:250, clone D1X5C, cat. no. 13333S, Cell Signaling Technologies, Inc.), mouse anti-Ki67 (1:2,000, clone 8D5, cat. no. 9449S, Cell Signaling Technologies, Inc.), goat anti-mouse IgG (HRP-linked, 1:2,000, cat. no. AP124P, MilliporeSigma), and goat anti-rabbit IgG (HRP-linked, 1:2,000, cat. no. AP132P, MilliporeSigma).

Techniques:

Journal: bioRxiv

Article Title: Tumor-specific neutrophils originating from meninges promote glioblastoma

doi: 10.1101/2023.05.23.542010

Figure Lengend Snippet: (A to G) Identification of tumor-specific neutrophils (TSNs) in human glioblastomas (GBM). (A) Diagram of profiling myeloid cells in human GBM tumors. (B) Major cell types identified in human GBM tumors. Baso, basophils; Ma_MRC1, Ma_BNIP3, Ma_LYZ, Ma_MKI67, macrophages with corresponding markers; Mi, microglia; N, neurons; Neu, neutrophils; NK, natural killer cells; Ol, oligodendrocytes; TSN, tumor-specific neutrophils; Tu, tumor cells. (C) Neutrophil subtypes are defined in human GBM tumors and patients’ blood. Neu_CXCR2, Neu_ISG15, Neu_S100A12, neutrophils with corresponding markers; TSN, tumor-specific neutrophils. (D) Top expressing genes in neutrophil subtypes as defined in (C) . (E) Violin plot of the expression of neutrophil marker genes ( CXCR2 and S100A8 ), immunosuppressive genes ( CD274 and IDO1 ), and myeloid recruitment-related genes ( CXCL8 , CCL3 , CCL4 , and IL1B ) in neutrophil subtypes. (F and G) Representative images of CD66b/CD274 immunofluorescence staining (F) or CD274 immunohistochemistry (G) of human GBM tumors. White arrows, CD66b + CD274 + cells; black arrow, CD274 + neutrophil with the lobulated nucleus. (H to L) TSNs are correlated with glioma grades and poor prognosis. (H) TSN scores are positively correlated with glioma grades in the TCGA dataset (Student’s t -test). (I) Correlation between TSN scores and CD274 expression levels in the TCGA dataset. The correlation coefficient (R) and p -value were calculated by Pearson correlation. (J) TSN scores are enriched in the mesenchymal subtype than in other molecular subtypes of human GBM (Student’s t -test). (K and L) Overall survival of TCGA GBM cases (K) or TCGA glioma cases (L) stratified by TSN scores (log-rank test).

Article Snippet: For the immunohistochemistry staining, 5 μm-thick paraffin sections were deparaffinized and stained with anti-human KI67 (1:500, TA802544, Origene), anti-mouse Ki67 (1:800, #12202, Cell Signaling Technology), anti-mouse Ly6G (1:200, #87048, Cell Signaling Technology), anti-mouse Cd8a (1:800, #98941, Cell Signaling Technology), or anti-mouse Cd274 (1:200, #64988, Cell Signaling Technology).

Techniques: Expressing, Marker, Immunofluorescence, Staining, Immunohistochemistry

Journal: bioRxiv

Article Title: Tumor-specific neutrophils originating from meninges promote glioblastoma

doi: 10.1101/2023.05.23.542010

Figure Lengend Snippet: (A) Dot plot of major cell types identified by profiling myeloid cells in human GBM tumors. Three top marker genes for each cell type are shown. Baso, basophils; Ma_MRC1, Ma_BNIP3, Ma_LYZ, Ma_MKI67, macrophages with corresponding markers; Mi, microglia; N, neurons; Neu, neutrophils; NK, natural killer cells; Ol, oligodendrocytes; TSN, tumor-specific neutrophils; Tu, tumor cells. (B) Expression of the neutrophil marker gene ( CXCR2 ) and immunosuppressive genes ( CD274 , IDO1, IDO2 , and CEACAM1 ) in major cell types identified in (A) . (C) Frequency of TSNs and other subtypes in the total neutrophils of GBM patients’ blood and tumors defined by scRNA-seq. (D) Representative image of CD66b/CD274 immunofluorescence staining of human low-grade glioma tumor. White arrows, CD66b + CD274 + cells.

Article Snippet: For the immunohistochemistry staining, 5 μm-thick paraffin sections were deparaffinized and stained with anti-human KI67 (1:500, TA802544, Origene), anti-mouse Ki67 (1:800, #12202, Cell Signaling Technology), anti-mouse Ly6G (1:200, #87048, Cell Signaling Technology), anti-mouse Cd8a (1:800, #98941, Cell Signaling Technology), or anti-mouse Cd274 (1:200, #64988, Cell Signaling Technology).

Techniques: Marker, Expressing, Immunofluorescence, Staining

Journal: bioRxiv

Article Title: Tumor-specific neutrophils originating from meninges promote glioblastoma

doi: 10.1101/2023.05.23.542010

Figure Lengend Snippet: (A) Summary of the generation and tumorigenicity of mouse LCPN and derived cells. (B) Representative images of Gfap, Ki67, Cd44, or Olig2 immunohistochemistry of intracranial LCPN glioma tumors in C57BL/6 wild-type mice. (C) Survival curves of C57BL/6 wild-type or nude mice intracranially implanted with LCPN cells (log-rank test). (D) mRNA levels of Y-antigen genes ( Uty , Kdm5d , and Peg3 ) in cultured LCPN and LCPNS cells were assessed by quantitative PCR analyses (Student’s t -test). ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: For the immunohistochemistry staining, 5 μm-thick paraffin sections were deparaffinized and stained with anti-human KI67 (1:500, TA802544, Origene), anti-mouse Ki67 (1:800, #12202, Cell Signaling Technology), anti-mouse Ly6G (1:200, #87048, Cell Signaling Technology), anti-mouse Cd8a (1:800, #98941, Cell Signaling Technology), or anti-mouse Cd274 (1:200, #64988, Cell Signaling Technology).

Techniques: Derivative Assay, Immunohistochemistry, Cell Culture, Real-time Polymerase Chain Reaction

Journal: bioRxiv

Article Title: Tumor-specific neutrophils originating from meninges promote glioblastoma

doi: 10.1101/2023.05.23.542010

Figure Lengend Snippet: (A and B) Differential expression of neutrophil recruitment-related chemokines in mouse glioma models. (A) Volcano plot comparing the bulk RNA-seq data of LCPN vs. GL261 tumor allografts intracranially implanted in C57BL/6 wild-type mice. Increased expression levels of Cxcl1 and Cxcl5 in LCPN tumors are highlighted. (B) mRNA levels of Cxcl1 and Cxcl5 in cultured GL261 and LCPN-SIIN cells were assessed by quantitative PCR analyses (Student’s t -test). (C to H) Cxcl1 deletion in glioma cells inhibits TSN recruitment and tumor growth. C57BL/6 wild-type mice were intracranially implanted with LCPNS-SIIN cells or LCPNS-SIIN-Cxcl1KO cells (clone #1 or clone #2). (C and D) The percentage of neutrophils (Cd45 + Cd11b + Ly6G + ) (C) or TSNs (Cd45 + Cd274 + Cd11b + Ly6G + ) (D) in total immune cells (Cd45 + ) of tumors were examined by FACS analyses (Student’s t -test). (E and F) Representative images (E) and quantification (F) of Cd8 immunohistochemistry of mouse glioma tumors (Student’s t -test). (G) The percentage of Pd1 + population in Cd8 + T cells of glioma tumors was quantified by FACS analyses (Student’s t -test). ( H ) Survival curves of the mice (log-rank test). (I to O) Cxcr2 deletion in host mice blocks neutrophil recruitment and tumor progression. Cxcr2 -/- or control Cxcr2 +/+ littermates were intracranially implanted with LCPNS-SIIN cells. (I) The percentage of neutrophils (Cd45 + Cd11b + Ly6G + ) in total immune cells (Cd45 + ) of tumors were examined by FACS analyses (Student’s t -test). (J and K) Representative images ( J ) and quantification (K) of Cd8 immunohistochemistry of mouse glioma tumors. (L and M) Representative images (L) and quantification (M) of Ki67 immunohistochemistry of glioma tumors. (N and O) Gross appearance (N) and tissue weights (O) of tumors in Cxcr2 -/- or control Cxcr2 +/+ mice (Student’s t -test). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: For the immunohistochemistry staining, 5 μm-thick paraffin sections were deparaffinized and stained with anti-human KI67 (1:500, TA802544, Origene), anti-mouse Ki67 (1:800, #12202, Cell Signaling Technology), anti-mouse Ly6G (1:200, #87048, Cell Signaling Technology), anti-mouse Cd8a (1:800, #98941, Cell Signaling Technology), or anti-mouse Cd274 (1:200, #64988, Cell Signaling Technology).

Techniques: Expressing, RNA Sequencing Assay, Cell Culture, Real-time Polymerase Chain Reaction, Immunohistochemistry

Journal: bioRxiv

Article Title: Tumor-specific neutrophils originating from meninges promote glioblastoma

doi: 10.1101/2023.05.23.542010

Figure Lengend Snippet: (A to D) C57BL/6 wild-type mice were intracranially implanted with glioma cells. (A) TSNs and other neutrophil subtypes identified in glioma tumors and the meninges and spleens of control or tumor-bearing mice. Neu_Camp, Neu_Cxcr2, Neu_Isg15, Neu_Mki67, Neu_S100a6, Neu_Slpi, neutrophils with corresponding markers; TSN, tumor-specific neutrophils. (B) Violin plot of the expression of the proliferation marker gene ( Mki67 ), immature neutrophil marker genes ( Cd177 and Camp ), neutrophil-derived proteolytic enzymes ( Mmp8 and Mmp9 ), neutrophil marker genes ( S100a8 and Cxcr2 ), interferon-stimulated gene ( Isg15 ), and immunosuppressive gene ( Cd274 ) in neutrophil subtypes. (C) Pseudotime trajectory analysis of neutrophil subtypes defined in (A) . (D) Presence of TSNs and other neutrophil subtypes in the spleens of control or tumor-bearing (TB) mice (upper panels), meninges of control or tumor-bearing mice, and glioma tumors (lower panels). ( E and F ) Parabiosis in mouse glioma models. (E) Diagram of the parabiosis of C57BL/6 wild-type mice with H11-tdTomato reporter mice followed by the intracranial implantation of glioma cells in C57BL/6 mice. (F) The percentage of tdTomato + cells in total immune cells (Cd45 + ) of the bone marrow, meninges, spleens, and blood of C57BL/6 mice after parabiosis. n.s., not significant.

Article Snippet: For the immunohistochemistry staining, 5 μm-thick paraffin sections were deparaffinized and stained with anti-human KI67 (1:500, TA802544, Origene), anti-mouse Ki67 (1:800, #12202, Cell Signaling Technology), anti-mouse Ly6G (1:200, #87048, Cell Signaling Technology), anti-mouse Cd8a (1:800, #98941, Cell Signaling Technology), or anti-mouse Cd274 (1:200, #64988, Cell Signaling Technology).

Techniques: Expressing, Marker, Derivative Assay

Journal: bioRxiv

Article Title: Tumor-specific neutrophils originating from meninges promote glioblastoma

doi: 10.1101/2023.05.23.542010

Figure Lengend Snippet: (A to C) Presence of TSNs in the meninges of mouse glioma models. C57BL/6 wild-type mice were intracranially implanted with glioma cells. (A) TSNs and other neutrophil subtypes identified in mouse meninges and glioma tumors. Neu_Camp, Neu_Cxcr2, Neu_Isg15, Neu_Mki67, Neu_S100a6, Neu_Slpi, neutrophils with corresponding markers; TSN, tumor-specific neutrophils. The black arrow denotes TSNs in the mouse meninges of tumor-bearing condition. (B) TSN frequency in the total neutrophils of mouse spleens and meninges defined by scRNA-seq. (C) The percentage of TSNs (Cd45 + Cd274 + Cd11b + Ly6G + ) in neutrophils (Cd45 + Cd11b + Ly6G + ) of the spleens, meninges, and gliomas of tumor-bearing mice was quantified by FACS analyses (Student’s t -test). ( D to G ) Tumor-induced myelopoiesis in the meninges. C57BL/6 wild-type mice were intracranially implanted with glioma cells. (D) The percentage of neutrophils (Cd45 + Cd11b + Ly6G + ) in total immune cells (Cd45 + ) of tumors at different time points post-implantation. (E) The percentage of granulocyte-monocyte progenitors (GMP, Lin - Sca-1 - c-Kit + Cd34 + Cd16/32 + ; Lin: B220, Cd2, Cd3, Cd5, Cd8, Gr-1, and Ter119) in total Lin - cells of the bone marrow, meninges, spleens, and gliomas of tumor-bearing mice were determined at day 5 post-implantation by FACS analyses (Student’s t -test). (F) Neutrophil proliferation occurs in the meninges. Ki67-RFP reporter mice were intracranially implanted with glioma cells. The percentage of Ki67-RFP + cells in neutrophils (Cd45 + Cd11b + Ly6G + ) of the spleens, meninges, and tumors were assessed by FACS analyses (Student’s t -test). ( G ) Cxcr2 deletion in host mice retains neutrophils in the meninges. Cxcr2 -/- or control Cxcr2 +/+ mice were intracranially implanted with glioma cells. The percentage of neutrophils (Cd45 + Cd11b + Ly6G + ) in total immune cells (Cd45 + ) of the meninges was quantified by FACS analyses (Student’s t -test). (H) Peripheral circulating immune cells were not sufficient to count for TSNs in mouse gliomas. A parabiosis between C57BL/6 wild-type mice and H11-tdTomato reporter mice was established. C57BL/6 mice in parabiotic pairs were then intracranially implanted with glioma cells. The percentage of tdTomato + cells in neutrophils (Cd45 + Cd11b + Ly6G + ) of the bone marrow, meninges, spleens, and tumors of C57BL/6 mice was determined by FACS analyses (Student’s t -test). ( I to K ) Presence of TSNs in the meninges of human GBM patients. ( I ) Major cell types identified in GBM patients’ meninges. B, B cells; DC, dendritic cells; En, endothelial cells; Flc, fibroblast-like cells; Ma, macrophages; Mast, mast cells; Mo, monocytes; N, neurons; Neu, neutrophils; SM, smooth muscle cells; T/NK, T cells / natural killer cells. (J) TSN frequency in the total neutrophils of GBM patients’ blood and meninges defined by scRNA-seq. (K) The percentage of neutrophils (CD45 + CD11b + CD14 - CD15 + HLA-DR - CD33 low CD66b + ) in total immune cells (CD45 + ) of the meninges of GBM or control patients was examined by FACS analyses (Student’s t -test). (L and M) Regulation of human TSN development by distinct signaling pathways. (L) KEGG enrichment diagram of human TSN marker genes. (M) Heat map of SCENIC binary regulon activities in TSNs and other neutrophil subtypes. Components of the NF-κB pathway are highlighted in orange. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: For the immunohistochemistry staining, 5 μm-thick paraffin sections were deparaffinized and stained with anti-human KI67 (1:500, TA802544, Origene), anti-mouse Ki67 (1:800, #12202, Cell Signaling Technology), anti-mouse Ly6G (1:200, #87048, Cell Signaling Technology), anti-mouse Cd8a (1:800, #98941, Cell Signaling Technology), or anti-mouse Cd274 (1:200, #64988, Cell Signaling Technology).

Techniques: Marker

Journal: Scientific Reports

Article Title: Cilia-associated wound repair mediated by IFT88 in retinal pigment epithelium

doi: 10.1038/s41598-023-35099-3

Figure Lengend Snippet: Defective cilia promote RPE wound healing by controlling proliferation. ( A ) Immunofluorescence analysis of phalloidin (red) and Ki67 (yellow) antibody in RPE flatmounts of RPE-cKO and control mice at 24 h and 48 h post laser treatment. RPE flatmounts were prepared from adult RPE-cKO mice and control mice 24 h and 48 h after laser photocoagulation (24 h: n = 6 and 10 eyes; 48 h: n = 4 and 8 eyes) and stained with Ki67 (proliferative cell marker) and phalloidin (cell border). ( B ) Quantification of wound size in RPE-cKO and control groups based on phalloidin staining, which was analyzed by ImageJ with manual correction (blinded to sample groups during analysis). ( C ) Quantification of cell number within laser area (200 μm). ( D ) Quantification of Ki67-positive cell number within 500 μm. Scale bars: 20 μm. Statistical analyses were performed using Student’s t-test, p < 0.05 was considered statistically significant.

Article Snippet: Primary antibodies, source, catalog number and dilutions are as follows: anti-Arl13b mouse (Antibodies Incorporated, N295B/66, 1:2000); anti-zonula occludens-1 (ZO-1) rabbit (Invitrogen, 617300, 1:350); anti-Cre mouse (Millipore, clone 2D8, 1:400); anti-RPE65 mouse (Abcam, ab13826, 1:500); anti-Ezrin mouse (Millipore, E8897, 1:500); anti-Ki67 mouse (Cell Signaling, 9449 T, 1:500); rhodamine phalloidin (Invitrogen, R415, 1:1000).

Techniques: Immunofluorescence, Staining, Marker

Journal: Brain

Article Title: Predicting progression to Alzheimer’s disease with human hippocampal progenitors exposed to serum

doi: 10.1093/brain/awac472

Figure Lengend Snippet: Outline of the experimental design and sample collection . ( A ) HN is regulated by a complex microenvironment, composed of blood vessels and various cell types such as hippocampal neural stem cells (NSCs), neural progenitor cells (NPCs), neuroblasts, immature/mature granule cells (i.e. neurons), microglia and astrocytes (i.e. neurogenic niche). Blood-derived factors, delivered to the niche by its rich vasculature, play a fundamental role in modulating HN. We aimed to model the role of systemic environment on the hippocampal neurogenic process during Alzheimer’s disease progression, by treating a human HPC line with 1% longitudinal serum at different stages of HN (i.e. proliferation and differentiation of HPCs). ( B ) Longitudinal serum samples were collected during annual follow-up visits from 56 participants diagnosed with MCI at baseline ( n = 38 converted to Alzheimer’s disease, n = 18 remained cognitively stable). A total of 338 samples were analysed. For each sample, three biological replicates (cells of three different passage numbers) were used and for each biological replicate, there were technical triplicates. ( C ) Neurogenic markers measured in the proliferation and differentiation phases of the assay are outlined ( top ) and representative images of cells positive for Ki67, CC3, Nestin, Sox2, MAP2 and DCX are shown ( bottom ). Scale bar = 100 μm. ( D ) An overview of the proliferation and differentiation phases in the assay. HPC0A07/03C cell line was treated with 1% serum samples from MCI converters and non-converters collected at sequential follow-up visits. Proliferation medium included EGF, bFGF and 4-OHT. Differentiation medium lacked these factors. To analyse serum effects on proliferation, 24 h after seeding, medium was replaced with proliferation medium supplemented with 1% serum. Cells were fixed 48 h later and subjected to ICC. To analyse the effects of serum on differentiation, at the end of proliferation phase, medium was replaced with differentiation medium supplemented with 1% serum. Cells were fixed 7 days later and subjected to ICC. c = converters; nc = non-converters. Panel A was created with BioRender.com.

Article Snippet: Mouse monoclonal anti-Ki67 (Cell Signaling, #9449, 1:800) was used to assess proliferation (i.e. HPCs in active phases of the cell cycle such as G1, S, G2 and mitosis); rabbit monoclonal anti-CC3 (Cell Signaling, #9664, 1:500) to assess apoptotic cell death; mouse monoclonal anti-Nestin clone 10C2 (Sigma Aldrich, #MAB5326, 1:1000) and rabbit polyclonal anti-Sox2 (SRY-Box Transcription Factor 2) (Sigma Aldrich, #AB5603, 1:1000) to assess neural stemcellness; rabbit polyclonal anti-DCX (Abcam, #ab18723, 1:500) for neuroblasts and mouse monoclonal anti-MAP2 (Abcam, #ab11267, 1:500) for mature neurons.

Techniques: Derivative Assay

Journal: Brain

Article Title: Predicting progression to Alzheimer’s disease with human hippocampal progenitors exposed to serum

doi: 10.1093/brain/awac472

Figure Lengend Snippet: Exposure to 1% serum from MCI converters leads to decreased proliferation, increased cell death and increased neurogenesis . ( A ) Representative images of proliferation phase cells treated with serum from the same individual. Left (MCI panel): serum sample from 1 year before conversion. Right (AD panel): serum sample taken at the time of conversion to Alzheimer’s disease (AD). Nuclei are stained with DAPI. Ki67 and CC3 were used to label proliferating and apoptotic cells, respectively. ( B – D ) Modelled trajectories (with 95% CIs) of linear mixed-effects regression models fitted to the proliferation phase data. Time of conversion to Alzheimer’s disease was assigned 0, and the number of years before conversion were assigned negative values (i.e. 1 year before conversion is −1). Longitudinal serum samples from MCI converters increased average cell number ( B ), decreased proliferation (% Ki67 + ) ( C ) and increased apoptotic cell death (% CC3 + ) ( D ). Slopes (β coefficient estimates) are indicated within the plots. ( E ) Representative images of differentiation phase cells treated with serum from the same individual. Left (MCI panel): serum sample from 1 year before conversion. Right (AD panel): serum sample taken at the time of conversion to AD. Nuclei are stained with DAPI. DCX and MAP2 were used to label neuroblasts and mature neurons, respectively. ( F – H ) Modelled trajectories (with 95% CIs) of linear mixed-effects regression models fitted to the differentiation phase data. Time of conversion to Alzheimer’s disease was assigned 0, and the number of years before conversion were assigned negative values (i.e. 1 year before conversion is −1). Longitudinal serum samples from MCI converters increased average cell number ( F ), neuroblasts (% DCX + ) ( G ) and mature neurons (% MAP2 + ) ( H ). Slopes (β coefficient estimates) are indicated within the plots. Scale bar = 100 μm.

Article Snippet: Mouse monoclonal anti-Ki67 (Cell Signaling, #9449, 1:800) was used to assess proliferation (i.e. HPCs in active phases of the cell cycle such as G1, S, G2 and mitosis); rabbit monoclonal anti-CC3 (Cell Signaling, #9664, 1:500) to assess apoptotic cell death; mouse monoclonal anti-Nestin clone 10C2 (Sigma Aldrich, #MAB5326, 1:1000) and rabbit polyclonal anti-Sox2 (SRY-Box Transcription Factor 2) (Sigma Aldrich, #AB5603, 1:1000) to assess neural stemcellness; rabbit polyclonal anti-DCX (Abcam, #ab18723, 1:500) for neuroblasts and mouse monoclonal anti-MAP2 (Abcam, #ab11267, 1:500) for mature neurons.

Techniques: Staining

Journal: Brain

Article Title: Predicting progression to Alzheimer’s disease with human hippocampal progenitors exposed to serum

doi: 10.1093/brain/awac472

Figure Lengend Snippet: Exposure to 1% serum from MCI converters leads to differential changes in average cell number, proliferation and neuronal differentiation compared to non-converters . ( A and B ) Modelled trajectories (with 95% CIs) of linear mixed-effects regression models fitted to the proliferation phase data. Time to last visit (for non-converters) and time of conversion to Alzheimer’s disease (for converters) was assigned 0, and the number of years before that were assigned negative values (i.e. 1 year before conversion is −1). Longitudinal serum samples from MCI converters (turquoise) predicted overall higher average cell number ( A ) and proliferation (% Ki67 + ) ( B ) compared to non-converters (red). Slopes (β coefficient estimates) are indicated within the plots. ( C and D ) Modelled trajectories (with 95% CIs) of linear mixed-effects regression models fitted to the differentiation phase data. Time to last visit (for non-converters) and time of conversion to Alzheimer’s disease (for converters) was assigned 0, and the number of years before that were assigned negative values (i.e. 1 year before conversion is −1). Longitudinal serum samples from MCI converters (turquoise) predicted overall lower average cell number ( C ) and higher neuronal differentiation (% MAP2 + ) ( D ) compared to non-converters (red). Slopes (β coefficient estimates) are indicated within the plots.

Article Snippet: Mouse monoclonal anti-Ki67 (Cell Signaling, #9449, 1:800) was used to assess proliferation (i.e. HPCs in active phases of the cell cycle such as G1, S, G2 and mitosis); rabbit monoclonal anti-CC3 (Cell Signaling, #9664, 1:500) to assess apoptotic cell death; mouse monoclonal anti-Nestin clone 10C2 (Sigma Aldrich, #MAB5326, 1:1000) and rabbit polyclonal anti-Sox2 (SRY-Box Transcription Factor 2) (Sigma Aldrich, #AB5603, 1:1000) to assess neural stemcellness; rabbit polyclonal anti-DCX (Abcam, #ab18723, 1:500) for neuroblasts and mouse monoclonal anti-MAP2 (Abcam, #ab11267, 1:500) for mature neurons.

Techniques:

Journal: Brain

Article Title: Predicting progression to Alzheimer’s disease with human hippocampal progenitors exposed to serum

doi: 10.1093/brain/awac472

Figure Lengend Snippet: Predictors of progression to Alzheimer’s disease from stepwise logistic regression analysis

Article Snippet: Mouse monoclonal anti-Ki67 (Cell Signaling, #9449, 1:800) was used to assess proliferation (i.e. HPCs in active phases of the cell cycle such as G1, S, G2 and mitosis); rabbit monoclonal anti-CC3 (Cell Signaling, #9664, 1:500) to assess apoptotic cell death; mouse monoclonal anti-Nestin clone 10C2 (Sigma Aldrich, #MAB5326, 1:1000) and rabbit polyclonal anti-Sox2 (SRY-Box Transcription Factor 2) (Sigma Aldrich, #AB5603, 1:1000) to assess neural stemcellness; rabbit polyclonal anti-DCX (Abcam, #ab18723, 1:500) for neuroblasts and mouse monoclonal anti-MAP2 (Abcam, #ab11267, 1:500) for mature neurons.

Techniques:

Journal: Brain

Article Title: Predicting progression to Alzheimer’s disease with human hippocampal progenitors exposed to serum

doi: 10.1093/brain/awac472

Figure Lengend Snippet: Average cell number and Ki67 during proliferation, and CC3 during differentiation, combined with education in years can predict progression from MCI to Alzheimer’s disease . ( A ) ROC curve for the logistic regression model predicting progression from MCI to Alzheimer’s disease. AUC for the model, an indicator of the discriminative performance, is 0.967. Sensitivity = 92.1%, specificity = 94.1%, positive predictive value = 97.2% and negative predictive value = 84.2%. ( B ) ROC curves for each four individual predictors included in the full logistic regression model. ( C ) Odds ratios for the four predictors. Blue and red indicate >1 and <1, respectively. * P < 0.05. *** P < 0.001. ( D ) ROC curve for the cross-validated logistic regression model predicting progression to Alzheimer’s disease. Internal validation of the model was done with repeated k -fold cross-validation ( k = 5, 1000 repeats) using SMVs (radial basis function kernel). AUC = 0.93, sensitivity 90.3% and specificity 79.0%.

Article Snippet: Mouse monoclonal anti-Ki67 (Cell Signaling, #9449, 1:800) was used to assess proliferation (i.e. HPCs in active phases of the cell cycle such as G1, S, G2 and mitosis); rabbit monoclonal anti-CC3 (Cell Signaling, #9664, 1:500) to assess apoptotic cell death; mouse monoclonal anti-Nestin clone 10C2 (Sigma Aldrich, #MAB5326, 1:1000) and rabbit polyclonal anti-Sox2 (SRY-Box Transcription Factor 2) (Sigma Aldrich, #AB5603, 1:1000) to assess neural stemcellness; rabbit polyclonal anti-DCX (Abcam, #ab18723, 1:500) for neuroblasts and mouse monoclonal anti-MAP2 (Abcam, #ab11267, 1:500) for mature neurons.

Techniques:

Journal: Cell reports

Article Title: Deletion of STAT3 from Foxd1 cell population protects mice from kidney fibrosis by inhibiting pericytes trans-differentiation and migration

doi: 10.1016/j.celrep.2022.110473

Figure Lengend Snippet: KEY RESOURCES TABLE Primary and secondary antibodies

Article Snippet: LTL was purchased from Vector laboratories (cat. no. NC0127927). . Antibodies: Anti-pSTAT3 Tyrosine 705 (cat. no. 9145) and anti-STAT3 (cat. no. 9139), anti-Ki67 (cat. no. 9449), anti-F4/80 (cat. no. 30325), pH2AX (cat. no. 9718), pSMAD2 (cat. no. 18338) were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: