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Elabscience Biotechnology percp anti human cd8a antibody

BRD7 downregulated PD-L1 expression and enhanced the cytotoxicity of T lymphocytes against NPC cells. (A) CIBERSORT determined the proportion of immune cell populations in NPC. (B) <t>CD8</t> + T cell infiltration plays a vital role in NPC. (C) Correlation analysis between the expression of BRD7 and immune cells abundance. (D) Correlation analysis between BRD7 expression level and immune cell subtypes in GSE102349 of NPC. (E) Clonogenic assays of 5-8F and CNE2 cells stably transfected with BRD7 overexpression and knockdown or empty vector plasmids with or without T cells co-culture and PD-L1 antibody incubation. Atezolizumab: the PD-L1 antibody. (F) CCK-8 assay of 5-8F and CNE2 cells stably transfected with BRD7 overexpression and knockdown or empty vector plasmids with or without T cells co-culture and PD-L1 antibody incubation. Absorbance values were detected at 450 nm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant. (G) Flow cytometry detecting the apoptosis ratio of CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 overexpression or empty vector plasmids with or without PD-L1 antibody incubation. (H) Flow cytometry detecting the apoptosis ratio of CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 knockdown or empty vector plasmids. (I) Flow cytometry detecting the ratio of PD-1 + CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 overexpression or empty vector plasmids with or without PD-L1 antibody incubation. (J) Flow cytometry detecting the ratio of PD-1 + CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 knockdown or empty vector plasmids. (K) ELISA detecting the IFN-γ content in the culture medium of T cells co-cultured with 5-8F cells stably transfected with BRD7 overexpression or empty vector plasmids and CNE2 cells stably transfected with BRD7 knockdown or empty vector plasmids with or without PD-L1 antibody incubation for 24 hours. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.

Percp Anti Human Cd8a Antibody, supplied by Elabscience Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "BRD7 Inhibited Immune Escape in Nasopharyngeal Carcinoma via Inhibiting PD-L1 Expression"

Article Title: BRD7 Inhibited Immune Escape in Nasopharyngeal Carcinoma via Inhibiting PD-L1 Expression

Journal: International Journal of Biological Sciences

doi: 10.7150/ijbs.103703

Figure Legend Snippet: BRD7 downregulated PD-L1 expression and enhanced the cytotoxicity of T lymphocytes against NPC cells. (A) CIBERSORT determined the proportion of immune cell populations in NPC. (B) CD8 + T cell infiltration plays a vital role in NPC. (C) Correlation analysis between the expression of BRD7 and immune cells abundance. (D) Correlation analysis between BRD7 expression level and immune cell subtypes in GSE102349 of NPC. (E) Clonogenic assays of 5-8F and CNE2 cells stably transfected with BRD7 overexpression and knockdown or empty vector plasmids with or without T cells co-culture and PD-L1 antibody incubation. Atezolizumab: the PD-L1 antibody. (F) CCK-8 assay of 5-8F and CNE2 cells stably transfected with BRD7 overexpression and knockdown or empty vector plasmids with or without T cells co-culture and PD-L1 antibody incubation. Absorbance values were detected at 450 nm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant. (G) Flow cytometry detecting the apoptosis ratio of CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 overexpression or empty vector plasmids with or without PD-L1 antibody incubation. (H) Flow cytometry detecting the apoptosis ratio of CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 knockdown or empty vector plasmids. (I) Flow cytometry detecting the ratio of PD-1 + CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 overexpression or empty vector plasmids with or without PD-L1 antibody incubation. (J) Flow cytometry detecting the ratio of PD-1 + CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 knockdown or empty vector plasmids. (K) ELISA detecting the IFN-γ content in the culture medium of T cells co-cultured with 5-8F cells stably transfected with BRD7 overexpression or empty vector plasmids and CNE2 cells stably transfected with BRD7 knockdown or empty vector plasmids with or without PD-L1 antibody incubation for 24 hours. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.

Techniques Used: Expressing, Stable Transfection, Transfection, Over Expression, Knockdown, Plasmid Preparation, Co-Culture Assay, Incubation, CCK-8 Assay, Flow Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay

BRD7 could inhibit the immune escape of NPC. (A) Design of the experiment in vivo . (B) Images of the appearance of subcutaneous tumors in mice. n = 5 per group. (C) Statistical analysis of subcutaneous tumor weight. n = 5 per group. (D) Tumor growth curve. n = 5 per group. (E) Relative BRD7 and PD-L1 mRNA levels measured by q-PCR in tumor tissues. n = 5 per group. (F) Western blot analysis of BRD7, PD-L1, and PI3K/AKT pathway molecules in tumor tissues. (G) Representative images of immunohistochemical staining for BRD7, PD-L1, PI3K/AKT pathway molecules, and CD8 expression in tumor tissues. The scale bar is 20 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.

Figure Legend Snippet: BRD7 could inhibit the immune escape of NPC. (A) Design of the experiment in vivo . (B) Images of the appearance of subcutaneous tumors in mice. n = 5 per group. (C) Statistical analysis of subcutaneous tumor weight. n = 5 per group. (D) Tumor growth curve. n = 5 per group. (E) Relative BRD7 and PD-L1 mRNA levels measured by q-PCR in tumor tissues. n = 5 per group. (F) Western blot analysis of BRD7, PD-L1, and PI3K/AKT pathway molecules in tumor tissues. (G) Representative images of immunohistochemical staining for BRD7, PD-L1, PI3K/AKT pathway molecules, and CD8 expression in tumor tissues. The scale bar is 20 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.

Techniques Used: In Vivo, Western Blot, Immunohistochemical staining, Staining, Expressing

Image Search Results

Immunophenotyping of PBMCs from pregnant SARS-CoV-2 infected and recovered patients. (a) Malaysian Cohort 1: Description of PBMCs used for immunophenotyping collected from pregnant healthy controls (Preg-HC, blue), SARS-CoV-2 infected (Preg-INF, orange) and COVID-19 recovered (Preg-R, pink). (b) Unsupervised clustering of immune cells based on 14-color flow cytometry panel. Each PBMC sample from Preg-HC, Preg-INF, and Preg-R groups were concatenated bioinformatically using the FlowJo software. Two major clusters for lymphocytes (pink) and monocytes (blue) population were identified and presented on a UMAP plot. The combined cell population is shown in gray. (c) UMAP plot characterising 7 identified immune subsets; monocytes (cyan), CD4 + T cells (navy), CD8 + T cells (red), NKT cells (lilac), CD19 + B cells (green), NK cells (rose), and Tregs (yellow). (d) Overlay of UMAP to show the different immune cell landscape; Preg-HC (blue), Preg-INF (orange), and Preg-R (pink) patient groups. The combined cell population is shown in gray. (e) Delineation of all CD8 + T subsets. UMAP overlay showing the central memory (blue), early (red), intermediate (orange), late effector (bright green), and Naïve (dark green) CD8 + T cells. The combined cell population is shown in rose. (f) UMAP overlay depicting the various CD8 + T cells based on the different patient groups. Preg-HC (cyan), Preg-R (pink), and Preg-D (orange). The combined cell population is shown in gray. (g) Original FACS plots of cytotoxic CD8 + T cells. Identification of Naïve, memory, and effector memory CD8 + T cells based on CD45RA and CCR7 markers. FACS plots show the CD45RA + CCR7 + naïve, CD45RA - CCR7 + CM cells, CD45RA +high CCR7 - late EM, CD45RA +mid CCR7 + intermediate EM, CD45RA - CCR7 - early EM cells in the different patient groups. (h) The percentage of Naïve (upper left), intermediate (EM II, right) and late EM III (bottom left) cells are shown as violin plots with each dot represents an individual sample. P-values show the significance among Preg-HC, Preg-INF and Preg-R groups and compared using Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test. P ≤0.05 considered significant (*p ≤0.05).

Journal: Frontiers in Immunology

Article Title: Single-cell RNA-sequencing highlights a curtailed NK cell function in convalescent COVID-19 pregnant women

doi: 10.3389/fimmu.2025.1560391

Figure Lengend Snippet: Immunophenotyping of PBMCs from pregnant SARS-CoV-2 infected and recovered patients. (a) Malaysian Cohort 1: Description of PBMCs used for immunophenotyping collected from pregnant healthy controls (Preg-HC, blue), SARS-CoV-2 infected (Preg-INF, orange) and COVID-19 recovered (Preg-R, pink). (b) Unsupervised clustering of immune cells based on 14-color flow cytometry panel. Each PBMC sample from Preg-HC, Preg-INF, and Preg-R groups were concatenated bioinformatically using the FlowJo software. Two major clusters for lymphocytes (pink) and monocytes (blue) population were identified and presented on a UMAP plot. The combined cell population is shown in gray. (c) UMAP plot characterising 7 identified immune subsets; monocytes (cyan), CD4 + T cells (navy), CD8 + T cells (red), NKT cells (lilac), CD19 + B cells (green), NK cells (rose), and Tregs (yellow). (d) Overlay of UMAP to show the different immune cell landscape; Preg-HC (blue), Preg-INF (orange), and Preg-R (pink) patient groups. The combined cell population is shown in gray. (e) Delineation of all CD8 + T subsets. UMAP overlay showing the central memory (blue), early (red), intermediate (orange), late effector (bright green), and Naïve (dark green) CD8 + T cells. The combined cell population is shown in rose. (f) UMAP overlay depicting the various CD8 + T cells based on the different patient groups. Preg-HC (cyan), Preg-R (pink), and Preg-D (orange). The combined cell population is shown in gray. (g) Original FACS plots of cytotoxic CD8 + T cells. Identification of Naïve, memory, and effector memory CD8 + T cells based on CD45RA and CCR7 markers. FACS plots show the CD45RA + CCR7 + naïve, CD45RA - CCR7 + CM cells, CD45RA +high CCR7 - late EM, CD45RA +mid CCR7 + intermediate EM, CD45RA - CCR7 - early EM cells in the different patient groups. (h) The percentage of Naïve (upper left), intermediate (EM II, right) and late EM III (bottom left) cells are shown as violin plots with each dot represents an individual sample. P-values show the significance among Preg-HC, Preg-INF and Preg-R groups and compared using Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test. P ≤0.05 considered significant (*p ≤0.05).

Article Snippet: Anti-Human CD8a PerCP-eFlourTM 710 (clone SK1) , Thermo Fisher Scientific (eBioscienceTM) , Cat#46-0087-42.

Techniques: Infection, Flow Cytometry, Software

Increased effector CD4 + T cells and NKT cells in infected and recovered pregnant women. (a) UMAP analysis of total CD4 + T cells into different CD4 + T subsets including Naïve T cells (green), central memory T cells (CM; blue), early effector T cells (EMI; red), late effector cells (EMII; orange), and Tregs (yellow). The combined cell population is shown in gray. (b) UMAP for different CD4 + T subsets showing Preg-HC (cyan), Preg-R (pink), and Preg-INF (orange). The combined cell population is shown in gray. (c) Original FACS plots identifies CD4 + T cells subsets based on CCR7 and CD45RA markers. Classification of naïve, memory and effector memory CD4 + T cells based on CD45RA and CCR7 markers. FACS plots show the CD45RA + CCR7 + Naïve, CD45RA - CCR7 + CM cells, CD45RA +high CCR7 - late EM, CD45RA +mid CCR7 + intermediate EM, CD45RA - CCR7 - early EM cells. (d) The percentage of Naïve, early, and late EM cells are shown in violin plots. P-values show the significance among Preg-HC, Preg-INF, and Preg-R groups and compared using Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test. P ≤0.05 considered significant (*p ≤0.05). (e) UMAP plots showing the distribution of NKT cells (blue) in the CD3 + T cell compartment. (f) UMAP displaying the overlay with different patient groups. Preg-HC (cyan), Preg-R (pink), and Preg-INF (orange) for NKT cells. Combined samples are shown in gray. (g) Original FACS plots presenting the expression of CD56 on CD3 + CD8 + T cells with antibody markers staining for CD8 and CD56. (h) The percentage of NKT cells displayed by violin plots among Preg-HC, Preg-INF, and Preg-R groups. Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test was used to determine significance (*p ≤0.05).

Journal: Frontiers in Immunology

Article Title: Single-cell RNA-sequencing highlights a curtailed NK cell function in convalescent COVID-19 pregnant women

doi: 10.3389/fimmu.2025.1560391

Figure Lengend Snippet: Increased effector CD4 + T cells and NKT cells in infected and recovered pregnant women. (a) UMAP analysis of total CD4 + T cells into different CD4 + T subsets including Naïve T cells (green), central memory T cells (CM; blue), early effector T cells (EMI; red), late effector cells (EMII; orange), and Tregs (yellow). The combined cell population is shown in gray. (b) UMAP for different CD4 + T subsets showing Preg-HC (cyan), Preg-R (pink), and Preg-INF (orange). The combined cell population is shown in gray. (c) Original FACS plots identifies CD4 + T cells subsets based on CCR7 and CD45RA markers. Classification of naïve, memory and effector memory CD4 + T cells based on CD45RA and CCR7 markers. FACS plots show the CD45RA + CCR7 + Naïve, CD45RA - CCR7 + CM cells, CD45RA +high CCR7 - late EM, CD45RA +mid CCR7 + intermediate EM, CD45RA - CCR7 - early EM cells. (d) The percentage of Naïve, early, and late EM cells are shown in violin plots. P-values show the significance among Preg-HC, Preg-INF, and Preg-R groups and compared using Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test. P ≤0.05 considered significant (*p ≤0.05). (e) UMAP plots showing the distribution of NKT cells (blue) in the CD3 + T cell compartment. (f) UMAP displaying the overlay with different patient groups. Preg-HC (cyan), Preg-R (pink), and Preg-INF (orange) for NKT cells. Combined samples are shown in gray. (g) Original FACS plots presenting the expression of CD56 on CD3 + CD8 + T cells with antibody markers staining for CD8 and CD56. (h) The percentage of NKT cells displayed by violin plots among Preg-HC, Preg-INF, and Preg-R groups. Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test was used to determine significance (*p ≤0.05).

Article Snippet: Anti-Human CD8a PerCP-eFlourTM 710 (clone SK1) , Thermo Fisher Scientific (eBioscienceTM) , Cat#46-0087-42.

Techniques: Infection, Expressing, Staining

Validation of immunophenotyping studies and dysregulated humoral immune response in German cohort 2. (a) German Cohort 2: description of cohort and experimental plan using matched PBMCs and serum from pregnant healthy controls (Preg-HC) and COVID-19 recovered (Preg-R) pregnant women. (b) Reduced percentage of CD14 + monocytes and significantly increased CD14 + CD16 + in Preg-R patients. Box plots show median, interquartile range (IQR), and the whiskers corresponding to the highest and lowest points within 1.5 times of IQR. Each dot represents an individual sample. Wilcoxon rank-sum test was used for p-value significance to compare pregnant healthy control (Preg-HC) and COVID-19 recovered (Preg-R). P ≤0.05 considered significant (*p ≤0.05). (c) Box and whisker plot representing the percentage of Naïve, CM, EM I, and EM II CD8 + T cells in Preg-HC and Preg-R patients. CD8 + Naïve and CD8 + CM T cells were lower in Preg-R. CD8 + EM I and CD8 + EM II T cells were significantly increased Preg-R. Wilcoxon rank-sum test was used for p-value significance. P ≤0.05 considered significant (*p ≤0.05). (d) Box and whisker plot representing NKT cells in Preg-R compared with Preg-HC. (e) Box and whisker’s plot showing reduced CD56 + NK cells and CD56 + CD16 + NK II cells in Preg-R. Increased levels of CD56 - HLA-DR + lymphoid cells were significantly increased in Preg-R. Wilcoxon rank-sum test was used for p-value significance. P ≤0.05 considered significant (*p ≤0.05). (f) High levels of Nucleocapsid IgG antibody levels in serum of Preg-R group compared to Preg-HC (left graph). Wilcoxon rank-sum test was used for p-value significance. P ≤0.05 considered significant (*p ≤0.05). Spike S1 (middle graph) and RBD IgG concentration (ug/mL; right graph) at different collection time-points (CT) points (CT1, 0 day; CT2, 39 days; and CT3, 89 days post infection) during their pregnancy. Decreased levels of Spike S1 and RBD IgG antibodies 89 days post first collection time point in Preg-R group. Each dot represents an individual. Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test. P ≤0.05 considered significant (*p ≤0.05). (g) Examination of IL-10, MCP-1, and IL-8 levels in the serum of recovered women up to 89 days post infection. Each dot represents an individual on a box-whisker plot. CT1, 0 day; CT2, 39 days; and CT3, 89 days post infection-during their pregnancy Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test. P ≤0.05 considered significant (*p ≤0.05, **p ≤0.01, ***p ≤0.001, ****p ≤0.0001).

Journal: Frontiers in Immunology

Article Title: Single-cell RNA-sequencing highlights a curtailed NK cell function in convalescent COVID-19 pregnant women

doi: 10.3389/fimmu.2025.1560391

Figure Lengend Snippet: Validation of immunophenotyping studies and dysregulated humoral immune response in German cohort 2. (a) German Cohort 2: description of cohort and experimental plan using matched PBMCs and serum from pregnant healthy controls (Preg-HC) and COVID-19 recovered (Preg-R) pregnant women. (b) Reduced percentage of CD14 + monocytes and significantly increased CD14 + CD16 + in Preg-R patients. Box plots show median, interquartile range (IQR), and the whiskers corresponding to the highest and lowest points within 1.5 times of IQR. Each dot represents an individual sample. Wilcoxon rank-sum test was used for p-value significance to compare pregnant healthy control (Preg-HC) and COVID-19 recovered (Preg-R). P ≤0.05 considered significant (*p ≤0.05). (c) Box and whisker plot representing the percentage of Naïve, CM, EM I, and EM II CD8 + T cells in Preg-HC and Preg-R patients. CD8 + Naïve and CD8 + CM T cells were lower in Preg-R. CD8 + EM I and CD8 + EM II T cells were significantly increased Preg-R. Wilcoxon rank-sum test was used for p-value significance. P ≤0.05 considered significant (*p ≤0.05). (d) Box and whisker plot representing NKT cells in Preg-R compared with Preg-HC. (e) Box and whisker’s plot showing reduced CD56 + NK cells and CD56 + CD16 + NK II cells in Preg-R. Increased levels of CD56 - HLA-DR + lymphoid cells were significantly increased in Preg-R. Wilcoxon rank-sum test was used for p-value significance. P ≤0.05 considered significant (*p ≤0.05). (f) High levels of Nucleocapsid IgG antibody levels in serum of Preg-R group compared to Preg-HC (left graph). Wilcoxon rank-sum test was used for p-value significance. P ≤0.05 considered significant (*p ≤0.05). Spike S1 (middle graph) and RBD IgG concentration (ug/mL; right graph) at different collection time-points (CT) points (CT1, 0 day; CT2, 39 days; and CT3, 89 days post infection) during their pregnancy. Decreased levels of Spike S1 and RBD IgG antibodies 89 days post first collection time point in Preg-R group. Each dot represents an individual. Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test. P ≤0.05 considered significant (*p ≤0.05). (g) Examination of IL-10, MCP-1, and IL-8 levels in the serum of recovered women up to 89 days post infection. Each dot represents an individual on a box-whisker plot. CT1, 0 day; CT2, 39 days; and CT3, 89 days post infection-during their pregnancy Kruskal–Wallis test, adjusted for Dunn’s multiple comparisons test. P ≤0.05 considered significant (*p ≤0.05, **p ≤0.01, ***p ≤0.001, ****p ≤0.0001).

Article Snippet: Anti-Human CD8a PerCP-eFlourTM 710 (clone SK1) , Thermo Fisher Scientific (eBioscienceTM) , Cat#46-0087-42.

Techniques: Biomarker Discovery, Control, Whisker Assay, Concentration Assay, Infection

Single cell atlas and immune cell composition of healthy controls and recovered COVID-19 pregnant women. (a) Integrated UMAP (UMAP-CCA) of 30,394 cells derived from PBMCs. Sc-RNA-seq data were analyzed using Seurat pipeline. UMAP plots shown the different immune cell subsets based on distinct gene expression. Cell types are color-coded as shown in UMAP. (b) Dot plots show the expression level of canonical cell markers used to assign cell subset identification for major cell type including CD4 + T, CD8 + T, and B cells. (c) Feature plots show the key canonical markers used for identification of individual cell types. Green intensity shows increasing expression. (d) Percentage of major cell types in Preg-HC and Preg-R patients. (e) UMAP analysis of individual Preg-HC (blue) and Preg-R (orange) patient groups which highlight a reduced NK I cell population.

Journal: Frontiers in Immunology

Article Title: Single-cell RNA-sequencing highlights a curtailed NK cell function in convalescent COVID-19 pregnant women

doi: 10.3389/fimmu.2025.1560391

Figure Lengend Snippet: Single cell atlas and immune cell composition of healthy controls and recovered COVID-19 pregnant women. (a) Integrated UMAP (UMAP-CCA) of 30,394 cells derived from PBMCs. Sc-RNA-seq data were analyzed using Seurat pipeline. UMAP plots shown the different immune cell subsets based on distinct gene expression. Cell types are color-coded as shown in UMAP. (b) Dot plots show the expression level of canonical cell markers used to assign cell subset identification for major cell type including CD4 + T, CD8 + T, and B cells. (c) Feature plots show the key canonical markers used for identification of individual cell types. Green intensity shows increasing expression. (d) Percentage of major cell types in Preg-HC and Preg-R patients. (e) UMAP analysis of individual Preg-HC (blue) and Preg-R (orange) patient groups which highlight a reduced NK I cell population.

Article Snippet: Anti-Human CD8a PerCP-eFlourTM 710 (clone SK1) , Thermo Fisher Scientific (eBioscienceTM) , Cat#46-0087-42.

Techniques: Derivative Assay, RNA Sequencing, Gene Expression, Expressing

Reduced cytotoxic functions of NKT and NK cells in recovered pregnant women. (a) UMAP plots show the NKT (upper panel) and NK I (lower panel) cells from pregnant healthy controls and recovered patients. (b) Differential gene expression analysis of NKT, NK I, and CD8 + TEMRA cells using volcano plots. (c) GSEA pathway enrichment analysis for NK (I) Selected activated and suppressed pathways are shown on GSEA bubble plots. (d) KEGG pathway analysis of NK I cells. Most significantly pathways are shown on the GSEA plot. (e) Dot plot represents the selected cytotoxic function expressing genes in NKT (C3), NK I (C7), NK II (C8), and NK III (C18) cells. (f) Violin plots show significantly regulated genes related with cytotoxic functions in NK I cells. Each represents one cell. (g) Dot plot representing genes related with cytotoxic functions in CD8 + TEMRA (C2) cells.

Journal: Frontiers in Immunology

Article Title: Single-cell RNA-sequencing highlights a curtailed NK cell function in convalescent COVID-19 pregnant women

doi: 10.3389/fimmu.2025.1560391

Figure Lengend Snippet: Reduced cytotoxic functions of NKT and NK cells in recovered pregnant women. (a) UMAP plots show the NKT (upper panel) and NK I (lower panel) cells from pregnant healthy controls and recovered patients. (b) Differential gene expression analysis of NKT, NK I, and CD8 + TEMRA cells using volcano plots. (c) GSEA pathway enrichment analysis for NK (I) Selected activated and suppressed pathways are shown on GSEA bubble plots. (d) KEGG pathway analysis of NK I cells. Most significantly pathways are shown on the GSEA plot. (e) Dot plot represents the selected cytotoxic function expressing genes in NKT (C3), NK I (C7), NK II (C8), and NK III (C18) cells. (f) Violin plots show significantly regulated genes related with cytotoxic functions in NK I cells. Each represents one cell. (g) Dot plot representing genes related with cytotoxic functions in CD8 + TEMRA (C2) cells.

Article Snippet: Anti-Human CD8a PerCP-eFlourTM 710 (clone SK1) , Thermo Fisher Scientific (eBioscienceTM) , Cat#46-0087-42.

Techniques: Gene Expression, Expressing

Comparison of quality attributes of mesoCAR-T derived from cryopreserved and fresh PBMCs. ( A ) Proportions of CD3 + , CD3 + CD4 + , and CD3 + CD8 + cells post magnetic bead enrichment (n = 4). ( B ) Structure of the mesoCAR. Viability ( C ) and cell expansion potential ( D ) during mesoCAR-T manufacturing (n = 4). The histogram in ( C ) and ( D ) represent the viability and amplification of CAR-T cells on the 11th day of culture, respectively. ( E ) Percentage of CD3 + , CD4 + , CD8 + and mesoCAR + cells (n = 4). Comparative analysis of the differentiation states ( F ) and exhaustion ( G ) (n = 4). The histogram in E–G present the statistical analysis of the CAR-T product. ( H ) Toxicity capacity curves, tested at various E:T (4:1 or 2:1) against SKOV-3 cells within 24 h (n = 4), and the toxicity at 24 h was analyzed statistically. ( I ) Cytokine secretion (n = 4). The data presented in ( A ) includes both individual and average values, whereas ( C-I ) show median with 95% CI. Dots of the same shape represent data from one individual. The Kruskal–Wallis test ( A ) and the Friedman test ( C-I ) were used for statistical analysis, with a p < 0.05 was considered significant. * p < 0.05, “ns” indicates no significant difference.

Journal: Scientific Reports

Article Title: Comparative analysis and process optimization for manufacturing CAR-T using the PiggyBac system derived from cryopreserved versus fresh PBMCs

doi: 10.1038/s41598-025-89686-7

Figure Lengend Snippet: Comparison of quality attributes of mesoCAR-T derived from cryopreserved and fresh PBMCs. ( A ) Proportions of CD3 + , CD3 + CD4 + , and CD3 + CD8 + cells post magnetic bead enrichment (n = 4). ( B ) Structure of the mesoCAR. Viability ( C ) and cell expansion potential ( D ) during mesoCAR-T manufacturing (n = 4). The histogram in ( C ) and ( D ) represent the viability and amplification of CAR-T cells on the 11th day of culture, respectively. ( E ) Percentage of CD3 + , CD4 + , CD8 + and mesoCAR + cells (n = 4). Comparative analysis of the differentiation states ( F ) and exhaustion ( G ) (n = 4). The histogram in E–G present the statistical analysis of the CAR-T product. ( H ) Toxicity capacity curves, tested at various E:T (4:1 or 2:1) against SKOV-3 cells within 24 h (n = 4), and the toxicity at 24 h was analyzed statistically. ( I ) Cytokine secretion (n = 4). The data presented in ( A ) includes both individual and average values, whereas ( C-I ) show median with 95% CI. Dots of the same shape represent data from one individual. The Kruskal–Wallis test ( A ) and the Friedman test ( C-I ) were used for statistical analysis, with a p < 0.05 was considered significant. * p < 0.05, “ns” indicates no significant difference.

Article Snippet: PerCP-Cyanine5. 5 anti-human CD8a , Thermo Fisher Scientific, Waltham, Massachusetts , 45–0088-42.

Techniques: Comparison, Derivative Assay, Amplification

Optimized process for enhanced expansion and functionality of mesoCAR-T from cryopreserved PBMCs. ( A ) Optimization of electroporation density. ( B ) Optimization of culture density after electroporation. Viability ( C ) and expansion potential ( D ) during mesoCAR-T manufacturing. ( E ) Percentages of CD3 + , CD4 + , CD8 + , and mesoCAR + cells (n = 3). ( F ) Differentiation and exhaustion states (n = 3). ( G ) Proportions of apoptotic cells and CD3 + , CD4 + , CD8 + subsets expressing CD45RO - CD27 + (n = 3). ( H ) The secretion level of PD-1 at an E:T = 2:1 (n = 3-4). ( I ) Cytokine secretion (n = 3). ( J ) The cytotoxic capacity curves at different E:T ratios (4:1 or 2:1) against SKOV-3 cells, and the cytotoxicity at 4 h and 24 h is shown in the histogram. Data is presented as median with 95% CI. Each dot represents an individual. Statistical analysis was conducted using the linear regression ( A ), Kruskal–Wallis test ( C, D, J ) or Mann Whitney test ( B, E, F, G, H, I ), with a p value < 0.05 indicating significance. * p < 0.05, “ns” indicates no significant difference.

Journal: Scientific Reports

Article Title: Comparative analysis and process optimization for manufacturing CAR-T using the PiggyBac system derived from cryopreserved versus fresh PBMCs

doi: 10.1038/s41598-025-89686-7

Figure Lengend Snippet: Optimized process for enhanced expansion and functionality of mesoCAR-T from cryopreserved PBMCs. ( A ) Optimization of electroporation density. ( B ) Optimization of culture density after electroporation. Viability ( C ) and expansion potential ( D ) during mesoCAR-T manufacturing. ( E ) Percentages of CD3 + , CD4 + , CD8 + , and mesoCAR + cells (n = 3). ( F ) Differentiation and exhaustion states (n = 3). ( G ) Proportions of apoptotic cells and CD3 + , CD4 + , CD8 + subsets expressing CD45RO - CD27 + (n = 3). ( H ) The secretion level of PD-1 at an E:T = 2:1 (n = 3-4). ( I ) Cytokine secretion (n = 3). ( J ) The cytotoxic capacity curves at different E:T ratios (4:1 or 2:1) against SKOV-3 cells, and the cytotoxicity at 4 h and 24 h is shown in the histogram. Data is presented as median with 95% CI. Each dot represents an individual. Statistical analysis was conducted using the linear regression ( A ), Kruskal–Wallis test ( C, D, J ) or Mann Whitney test ( B, E, F, G, H, I ), with a p value < 0.05 indicating significance. * p < 0.05, “ns” indicates no significant difference.

Article Snippet: PerCP-Cyanine5. 5 anti-human CD8a , Thermo Fisher Scientific, Waltham, Massachusetts , 45–0088-42.

Techniques: Electroporation, Expressing, MANN-WHITNEY

The antibodies employed for staining and flow cytometric analysis of cell phenotypes.

Journal: Scientific Reports

Article Title: Comparative analysis and process optimization for manufacturing CAR-T using the PiggyBac system derived from cryopreserved versus fresh PBMCs

doi: 10.1038/s41598-025-89686-7

Figure Lengend Snippet: The antibodies employed for staining and flow cytometric analysis of cell phenotypes.

Article Snippet: PerCP-Cyanine5. 5 anti-human CD8a , Thermo Fisher Scientific, Waltham, Massachusetts , 45–0088-42.

Techniques: Staining

Journal: International Journal of Biological Sciences

Article Title: BRD7 Inhibited Immune Escape in Nasopharyngeal Carcinoma via Inhibiting PD-L1 Expression

doi: 10.7150/ijbs.103703

Figure Lengend Snippet: BRD7 downregulated PD-L1 expression and enhanced the cytotoxicity of T lymphocytes against NPC cells. (A) CIBERSORT determined the proportion of immune cell populations in NPC. (B) CD8 + T cell infiltration plays a vital role in NPC. (C) Correlation analysis between the expression of BRD7 and immune cells abundance. (D) Correlation analysis between BRD7 expression level and immune cell subtypes in GSE102349 of NPC. (E) Clonogenic assays of 5-8F and CNE2 cells stably transfected with BRD7 overexpression and knockdown or empty vector plasmids with or without T cells co-culture and PD-L1 antibody incubation. Atezolizumab: the PD-L1 antibody. (F) CCK-8 assay of 5-8F and CNE2 cells stably transfected with BRD7 overexpression and knockdown or empty vector plasmids with or without T cells co-culture and PD-L1 antibody incubation. Absorbance values were detected at 450 nm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant. (G) Flow cytometry detecting the apoptosis ratio of CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 overexpression or empty vector plasmids with or without PD-L1 antibody incubation. (H) Flow cytometry detecting the apoptosis ratio of CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 knockdown or empty vector plasmids. (I) Flow cytometry detecting the ratio of PD-1 + CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 overexpression or empty vector plasmids with or without PD-L1 antibody incubation. (J) Flow cytometry detecting the ratio of PD-1 + CD8 + T cells in T cells co-cultured with 5-8F or CNE2 cells stably transfected with BRD7 knockdown or empty vector plasmids. (K) ELISA detecting the IFN-γ content in the culture medium of T cells co-cultured with 5-8F cells stably transfected with BRD7 overexpression or empty vector plasmids and CNE2 cells stably transfected with BRD7 knockdown or empty vector plasmids with or without PD-L1 antibody incubation for 24 hours. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.

Article Snippet: T lymphocytes and tumor cells were cocultured at a ratio of 10:1 for 24 h. T cells were stained with an Annexin V-FITC/PI Apoptosis Detection Kit (Biosharp, China), APC Anti-Human CD3 Antibody (Elabscience, China), PerCP Anti-Human CD8a Antibody (Elabscience, China), and FITC Anti-Human CD279/PD-1 Antibody (Elabscience, China) according to the instructions.

Techniques: Expressing, Stable Transfection, Transfection, Over Expression, Knockdown, Plasmid Preparation, Co-Culture Assay, Incubation, CCK-8 Assay, Flow Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Biological Sciences

Article Title: BRD7 Inhibited Immune Escape in Nasopharyngeal Carcinoma via Inhibiting PD-L1 Expression

doi: 10.7150/ijbs.103703

Figure Lengend Snippet: BRD7 could inhibit the immune escape of NPC. (A) Design of the experiment in vivo . (B) Images of the appearance of subcutaneous tumors in mice. n = 5 per group. (C) Statistical analysis of subcutaneous tumor weight. n = 5 per group. (D) Tumor growth curve. n = 5 per group. (E) Relative BRD7 and PD-L1 mRNA levels measured by q-PCR in tumor tissues. n = 5 per group. (F) Western blot analysis of BRD7, PD-L1, and PI3K/AKT pathway molecules in tumor tissues. (G) Representative images of immunohistochemical staining for BRD7, PD-L1, PI3K/AKT pathway molecules, and CD8 expression in tumor tissues. The scale bar is 20 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.

Techniques: In Vivo, Western Blot, Immunohistochemical staining, Staining, Expressing

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