Journal: iScience

Article Title: ADAMTS18-fibronectin interaction regulates the morphology of liver sinusoidal endothelial cells

doi: 10.1016/j.isci.2024.110273

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-CD14 , Proteintech , Cat#17000-1-AP, RRID: AB_2919143.

Techniques: Recombinant, RNAscope, cDNA Synthesis, SYBR Green Assay, Modification, Staining, Peroxidation Assay, Immunoprecipitation, Activity Assay, Plasmid Preparation, Software

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: S100A9 + CD14 + monocytes contribute to anti-PD-1 immunotherapy resistance in advanced hepatocellular carcinoma by attenuating T cell-mediated antitumor function

doi: 10.1186/s13046-024-02985-1

Figure Lengend Snippet: Monocyte-related gene signature enriched in the PBMC of non-responders. a Schematic diagram of the study design and workflow. b UMAP visualization of 30,667 single-cell transcriptomes of immune cells identifying 23 populations and colored by cell type assignment. c Stacked histograms indicating the proportion of immune subsets in each patient. d Volcano plot depicting DEGs between NR and R. Four top genes enriched in the NR group are boxed with red dashed lines, and their expression is shown as UMAP feature plots ( f ). e GO pathway enrichment analysis of DEGs from PBMCs between NR and R. g UMAP visualization of monocytes. Left: Monocytes stratified by NR group (blue) and R group (yellow). Right: Monocytes stratified into four populations. h Proportions of monocyte subsets in each group. The numbers indicate the percentage of S100A9 + CD14 + monocytes

Article Snippet: Staining was carried out using a 1:200 dilution of rabbit polyclonal antibody for human S100A9 (Proteintech Cat# 26,992–1-AP; RRID: AB_2880716) and a 1:2000 dilution of rabbit polyclonal antibody against human CD14 (Proteintech Cat# 17,000–1-AP; RRID: AB_2074048), followed by incubation with secondary antibodies (Biolynx Cat# I20012C).

Techniques: Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: S100A9 + CD14 + monocytes contribute to anti-PD-1 immunotherapy resistance in advanced hepatocellular carcinoma by attenuating T cell-mediated antitumor function

doi: 10.1186/s13046-024-02985-1

Figure Lengend Snippet: Circulating S100A9 + CD14 + monocyte predicts unfavorable responses to anti-PD-1 immunotherapy. a UMAP visualization of immune cells between NR and R. S100A9 + CD14 + monocytes are boxed with red dashed lines. b Percentage of each immune subset between the NR group (blue) and R group (yellow), represented as box-whisker plots. Significance was evaluated using 2-way ANOVA. c Violin plots representing the percentage of S100A9 + CD14 + monocytes in each patient. d Survival curves showing overall survival stratified by the percentage of S100A9 + CD14 + monocyte. e Left: Representative image of dual immunofluorescence demonstrating S100A9 (red) and CD14 (green) positive cells and S100A9 + CD14 + monocytes (white triangles) in HCC tumors of a responder and a non-responder. Right: Number of S100A9 + CD14 + monocytes quantified in five randomly selected fields per patient ( n = 4 in NR; n = 4 in R). Scale bar, 50 μm. f Levels of plasma S100A9 in patients among different ICB best efficacy. g ROC curves and the area under the curve (AUC) of the plasma S100A9 level (red) or NLR (dashed blue) for discrimination between responders and non-responders. The cut-off value for S100A9 level was 308.110 ng/ml. h Best efficacy of patients stratified by plasma S100A9 levels. i Survival curves showing progress-free survival stratified by the plasma S100A9 level. P value in ( c and e ) were calculated using unpaired Student’s t-test. P values in ( d and i ) were calculated using the log-rank test

Article Snippet: Staining was carried out using a 1:200 dilution of rabbit polyclonal antibody for human S100A9 (Proteintech Cat# 26,992–1-AP; RRID: AB_2880716) and a 1:2000 dilution of rabbit polyclonal antibody against human CD14 (Proteintech Cat# 17,000–1-AP; RRID: AB_2074048), followed by incubation with secondary antibodies (Biolynx Cat# I20012C).

Techniques: Whisker Assay, Immunofluorescence, Clinical Proteomics

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: S100A9 + CD14 + monocytes contribute to anti-PD-1 immunotherapy resistance in advanced hepatocellular carcinoma by attenuating T cell-mediated antitumor function

doi: 10.1186/s13046-024-02985-1

Figure Lengend Snippet: Higher Mono_S100A9 signature score predicts worse ICB response and T cell dysfunction in cancer patients. a Genes in Mono_S100A9 signature. b Mono_S100A9 signature scores in patients with melanoma between NR and R. c Association of Mono_S100A9-signature score with overall survival within LIHC dataset obtained from TCGA. LIHC: Liver hepatocellular carcinoma. d Volcano plot showing DEGs of T cells between NR and R. e-g GSEA plots of the indicated signature genes from T cells between NR and R. h UMAP visualization of tumor infiltrating leukocytes (TILs) in HCC cohort SRP318499 or in HCC cohort CNP0000650 ( j ). i Heatmap plot showing the expression level of T-cell cytotoxicity genes in tumor-infiltrating T cells from HCC cohort SRP318499 and CNP0000650 ( k ). Patients were divided into two groups (Mono_CD14_S100A9 high (blue) and low (yellow)) based on their infiltration level of S100A9 + CD14 + monocytes

Article Snippet: Staining was carried out using a 1:200 dilution of rabbit polyclonal antibody for human S100A9 (Proteintech Cat# 26,992–1-AP; RRID: AB_2880716) and a 1:2000 dilution of rabbit polyclonal antibody against human CD14 (Proteintech Cat# 17,000–1-AP; RRID: AB_2074048), followed by incubation with secondary antibodies (Biolynx Cat# I20012C).

Techniques: Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: S100A9 + CD14 + monocytes contribute to anti-PD-1 immunotherapy resistance in advanced hepatocellular carcinoma by attenuating T cell-mediated antitumor function

doi: 10.1186/s13046-024-02985-1

Figure Lengend Snippet: Endogenous S100A9 enhances PD-L1 expression in monocytes to inhibit T-cell cytotoxicity. a KEGG pathway enrichment analysis of DEGs between S100A9 + CD14 + monocytes and other circulating cells. b Percentage of S100A9 + cells (left) and PD-L1 + cells (right) in PBMC from healthy donors (HD, n = 5) and patients with hepatocellular carcinoma (HCC, n = 7), colorectal cancer (CRC, n = 5), and biliary tract cancer (BTC, n = 7). c-e PD-L1 levels in S100A9 high or S100A9 low cells in CD14 + monocytes from patients with HCC, CRC, and BTC. f-h Correlation analysis between PD-L1 and S100A9 levels in CD14 + monocytes from patients with HCC, CRC, and BTC. i S100A9 and CD274 ( j ) mRNA levels of S100A9 -knockdown THP-1 cells transfected with si-S100A9#1, si- S100A9 #2, and si- S100A9 #3. k Representative flow cytometric plot (left) and quantification (right) of S100A9 levels in S100A9 -knockdown THP-1 cells transfected with sh- S100A9 #1 and sh- S100A9 #2. l Representative flow cytometric plot (left) and quantification (right) of PD-L1 levels in S100A9 -knockdown THP-1 cells transfected with sh- S100A9 #1 and sh- S100A9 #2. m the percentage of highly proliferated CFSE low of T cells co-cultured with S100A9 -knockdown THP-1 cells at 96 h ( n = 3). n TBX21 , PRF1 , IL2 , and GZMB mRNA levels of T cells after co-cultured with S100A9 -knockdown THP-1 cells for 24 h. P value in ( b ) was evaluated using the Mann–whitney U-test. P values in c-e were determined by two-tailed paired sample t-test. P values in i-m were determined by one-way ANOVA. P value in ( n ) was evaluated using 2-way ANOVA. Correlations were analyzed using the Spearman rank correlation test. * P < 0.05, ** P < 0·01, *** P < 0.001 and **** P < 0.0001

Article Snippet: Staining was carried out using a 1:200 dilution of rabbit polyclonal antibody for human S100A9 (Proteintech Cat# 26,992–1-AP; RRID: AB_2880716) and a 1:2000 dilution of rabbit polyclonal antibody against human CD14 (Proteintech Cat# 17,000–1-AP; RRID: AB_2074048), followed by incubation with secondary antibodies (Biolynx Cat# I20012C).

Techniques: Expressing, Knockdown, Transfection, Cell Culture, MANN-WHITNEY, Two Tailed Test

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: S100A9 + CD14 + monocytes contribute to anti-PD-1 immunotherapy resistance in advanced hepatocellular carcinoma by attenuating T cell-mediated antitumor function

doi: 10.1186/s13046-024-02985-1

Figure Lengend Snippet: Exogenous S100A9 enhances PD-L1 expression in monocytes to inhibit T-cell proliferation and cytotoxicity. a Representative flow cytometric plot (left) and quantification (right) of PD-L1 levels in primary human CD14 + monocytes ( n = 3) treated with PBS or rS100A9 for 8 h. b Similar to ( a ), PD-L1 levels in THP-1 cells treated with PBS ( n = 3), rS100A9 ( n = 3), or pre-incubated tasquinimod plus rS100A9 ( n = 3) for 24 h. c Schematic representation of the co-culture system. d Representative CFSE dilution profiles of T cells (left) and the percentage of highly proliferated CFSE low T cells at 96 h (right, n = 3). The peak of the CFSE-labelled unstimulated cells (gray, filled) is also shown. e The percentage of highly proliferated CFSE low T cells at 96 h in four groups: pre-incubated with IgG, pre-incubated with IgG and co-cultured with S100A9-treated THP-1, pre-incubated with αPD-1 antibody, pre-incubated with αPD-1 antibody and co-cultured with S100A9-treated THP-1. ( n = 3). f The RUNX3 and TBX21 mRNA levels in T cells co-cultured with THP-1 cells treated with PBS or rS100A9 for 24 h. g Schematic showing S100A9 binding sites on CD274 promoter (left). Luciferase activities relative to the control were shown on the right ( n = 3). MFI, mean of fluorescence intensity. CFSE: carboxyfluorescein succinimidyl ester. Unsti: unstimulated. Data are represented as mean ± S.E.M. P value in ( a ) was determined by two-tailed paired sample t-test. P value in ( d ) was determined using two-tailed unpaired Student’s t-tests. P value in ( b ) was determined by one-way ANOVA. P values in ( e-g) were evaluated using 2-way ANOVA. * P < 0.05, ** P < 0·01, *** P < 0.001 and **** P < 0.0001

Article Snippet: Staining was carried out using a 1:200 dilution of rabbit polyclonal antibody for human S100A9 (Proteintech Cat# 26,992–1-AP; RRID: AB_2880716) and a 1:2000 dilution of rabbit polyclonal antibody against human CD14 (Proteintech Cat# 17,000–1-AP; RRID: AB_2074048), followed by incubation with secondary antibodies (Biolynx Cat# I20012C).

Techniques: Expressing, Incubation, Co-Culture Assay, Cell Culture, Binding Assay, Luciferase, Control, Fluorescence, Two Tailed Test

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: S100A9 + CD14 + monocytes contribute to anti-PD-1 immunotherapy resistance in advanced hepatocellular carcinoma by attenuating T cell-mediated antitumor function

doi: 10.1186/s13046-024-02985-1

Figure Lengend Snippet: Working model for the effect of S100A9 + CD14 + monocytes contribute to anti-PD-1 immunotherapy resistance. Abundant blood S100A9 + CD14 + monocytes and high concentrate plasma S100A9 linked to poor HCC response to anti-PD-1. Mono_S100A9 signature inversely associated with survival of cancer patients. S100A9 enhanced PD-L1 expression on monocytes to inhibit T-cell proliferation and cytotoxicity. Blockage of S100A9 synergizes with anti-PD-1 drug to enhance HCC eradication

Article Snippet: Staining was carried out using a 1:200 dilution of rabbit polyclonal antibody for human S100A9 (Proteintech Cat# 26,992–1-AP; RRID: AB_2880716) and a 1:2000 dilution of rabbit polyclonal antibody against human CD14 (Proteintech Cat# 17,000–1-AP; RRID: AB_2074048), followed by incubation with secondary antibodies (Biolynx Cat# I20012C).

Techniques: Clinical Proteomics, Expressing

Journal: Neural Regeneration Research

Article Title: Genetic modification of miR-34a enhances efficacy of transplanted human dental pulp stem cells after ischemic stroke

doi: 10.4103/1673-5374.367831

Figure Lengend Snippet: Identification and characterization of hDPSCs. (A) Cell morphology of the isolated hDPSCs in the third passage (scale bar: 200 μm). (B) Expression of hDPSC surface antigens as determined by flow cytometry. Positive surface antigen: CD44; negative surface antigen: CD45 ( n = 3). (C) Representative images of immunofluorescent staining of hDPSC surface antigens. Positive surface antigens: CD90, CD105, CD44; negative surface antigens: CD14, CD34, CD45 ( n = 3) (scale bars: 50 μm). DAPI: 4′,6-Diamidino-2-phenylindole; hDPSCs: human dental pulp stem cells; P3: passage 3.

Article Snippet: The cells were then incubated with hDPSC markers, rabbit polyclonal anti-CD45 (1:200; Servicebio, Cat# GB113885), rabbit polyclonal anti-CD105 (1:200; Servicebio, Cat# GB113377), rabbit polyclonal anti-CD90 (1:200; Servicebio, Cat# GB11182), rabbit polyclonal anti-CD34 (1:200; Servicebio, Cat# GB111693), rabbit polyclonal anti-CD44 (1:200; Servicebio, Cat# GB113500), and rabbit polyclonal anti-CD14 antibody (1:200; Servicebio, Cat# GB11254) at 4°C overnight.

Techniques: Isolation, Expressing, Flow Cytometry, Staining