cd8 antibody  (Boster Bio)


Bioz Verified Symbol Boster Bio is a verified supplier
Bioz Manufacturer Symbol Boster Bio manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92

    Structured Review

    Boster Bio cd8 antibody
    Targeting USP7 can activate the p38 MAPK pathway to reprogram TAMs. ( A ) The strategy for sorting TAMs in TME by flow cytometry. ( B ) Heat maps illustrating the differentially expressed M1- and M2-related genes in TAMs between the P5091 group and the Control group based on the results of RNA sequencing. ( C ) RT-PCR further verifying the differentially expressed genes of sorted TAMs in each group. Data are presented as the mean ± SEM (n = 3). ( D ) KEGG analysis identifying 20 most obviously enriched pathways based on the differentially expressed genes of the two groups. ( E ) Western blotting detection of the expression of JNK, p-JNK, ERK1/2, p-ERK1/2, p38, p-p38, and β-actin in IL-4/13-BMDM M2 cells treated with P5091 (10 µM) at the indicated time points. ( F ) Flow cytometry analysis of CFSE expression on the surface of <t>CD8</t> + T cells in the presence of conditioned medium of IL-4/13-induced BMDM M2 cells from various indicated treatments. Treatments indicated: DMSO stimulation, P5091 (10 µM) stimulation, P5091 (10 µM) stimulation in the presence of inhibitors of p38 (SB203580, 10 µM), JNK (SP600125, 10 µM), Erk1/2 (U0126-EtOH, 10 µM). Data are presented as the mean ± SEM (n = 4).
    Cd8 Antibody, supplied by Boster Bio, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd8 antibody/product/Boster Bio
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd8 antibody - by Bioz Stars, 2022-10
    92/100 stars

    Images

    1) Product Images from "USP7 targeting modulates anti-tumor immune response by reprogramming Tumor-associated Macrophages in Lung Cancer"

    Article Title: USP7 targeting modulates anti-tumor immune response by reprogramming Tumor-associated Macrophages in Lung Cancer

    Journal: Theranostics

    doi: 10.7150/thno.47137

    Targeting USP7 can activate the p38 MAPK pathway to reprogram TAMs. ( A ) The strategy for sorting TAMs in TME by flow cytometry. ( B ) Heat maps illustrating the differentially expressed M1- and M2-related genes in TAMs between the P5091 group and the Control group based on the results of RNA sequencing. ( C ) RT-PCR further verifying the differentially expressed genes of sorted TAMs in each group. Data are presented as the mean ± SEM (n = 3). ( D ) KEGG analysis identifying 20 most obviously enriched pathways based on the differentially expressed genes of the two groups. ( E ) Western blotting detection of the expression of JNK, p-JNK, ERK1/2, p-ERK1/2, p38, p-p38, and β-actin in IL-4/13-BMDM M2 cells treated with P5091 (10 µM) at the indicated time points. ( F ) Flow cytometry analysis of CFSE expression on the surface of CD8 + T cells in the presence of conditioned medium of IL-4/13-induced BMDM M2 cells from various indicated treatments. Treatments indicated: DMSO stimulation, P5091 (10 µM) stimulation, P5091 (10 µM) stimulation in the presence of inhibitors of p38 (SB203580, 10 µM), JNK (SP600125, 10 µM), Erk1/2 (U0126-EtOH, 10 µM). Data are presented as the mean ± SEM (n = 4).
    Figure Legend Snippet: Targeting USP7 can activate the p38 MAPK pathway to reprogram TAMs. ( A ) The strategy for sorting TAMs in TME by flow cytometry. ( B ) Heat maps illustrating the differentially expressed M1- and M2-related genes in TAMs between the P5091 group and the Control group based on the results of RNA sequencing. ( C ) RT-PCR further verifying the differentially expressed genes of sorted TAMs in each group. Data are presented as the mean ± SEM (n = 3). ( D ) KEGG analysis identifying 20 most obviously enriched pathways based on the differentially expressed genes of the two groups. ( E ) Western blotting detection of the expression of JNK, p-JNK, ERK1/2, p-ERK1/2, p38, p-p38, and β-actin in IL-4/13-BMDM M2 cells treated with P5091 (10 µM) at the indicated time points. ( F ) Flow cytometry analysis of CFSE expression on the surface of CD8 + T cells in the presence of conditioned medium of IL-4/13-induced BMDM M2 cells from various indicated treatments. Treatments indicated: DMSO stimulation, P5091 (10 µM) stimulation, P5091 (10 µM) stimulation in the presence of inhibitors of p38 (SB203580, 10 µM), JNK (SP600125, 10 µM), Erk1/2 (U0126-EtOH, 10 µM). Data are presented as the mean ± SEM (n = 4).

    Techniques Used: Flow Cytometry, RNA Sequencing Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing

    Targeting USP7 inhibits tumor growth and induces local anti-tumor immunity in vivo . ( A ) Harvested and photographed tumors in the P5091 and the Control group. ( B ) Lewis tumor growth following P5091 or Vehicle treatment in vivo. Data are presented as the mean ± SEM (n = 6 per group). ( C ) Spider diagram of the tumor volume growth in each mouse from the P5091 group and the Control group. ( D ) Gating strategy for detection of the TAMs by flow cytometry. ( E-G ) Proportions of M1 ( E ) and M2 ( F ), and M1/M2 ratio ( G ) in the TME of the P5091 group and the Control group. ( H-M ) Percentages of MDSC ( H ), Treg ( I ), CD4 + T ( J ), CD8 + T ( K ), Th1 ( L ), and CTLs ( M ) within the TME in each group. Data are presented as the mean ± SEM (n = 7) for ( E-M ).
    Figure Legend Snippet: Targeting USP7 inhibits tumor growth and induces local anti-tumor immunity in vivo . ( A ) Harvested and photographed tumors in the P5091 and the Control group. ( B ) Lewis tumor growth following P5091 or Vehicle treatment in vivo. Data are presented as the mean ± SEM (n = 6 per group). ( C ) Spider diagram of the tumor volume growth in each mouse from the P5091 group and the Control group. ( D ) Gating strategy for detection of the TAMs by flow cytometry. ( E-G ) Proportions of M1 ( E ) and M2 ( F ), and M1/M2 ratio ( G ) in the TME of the P5091 group and the Control group. ( H-M ) Percentages of MDSC ( H ), Treg ( I ), CD4 + T ( J ), CD8 + T ( K ), Th1 ( L ), and CTLs ( M ) within the TME in each group. Data are presented as the mean ± SEM (n = 7) for ( E-M ).

    Techniques Used: In Vivo, Flow Cytometry

    Targeting USP7 activates local and systemic anti-tumor immunity. ( A ) Multicolor immunofluorescence detection of TAMs and CTLs in the TME of the P5091 group and the Control group. ( B-F ) Cytokines IFN-γ ( B ), TNF-α ( C ), IL-2 ( D ), IL-5 ( E ), and IL-6 ( F ) in the TME of each group were detected by Mul-Analyte Flow Assay Kit. ( G-K ) Proportion of Treg ( G ), CD4 + T ( H ), CD8 + T ( I ), Th1 ( J ), and CTLs ( K ) in the spleen of each group. Data are presented as the mean ± SEM (n = 7) for ( B-K ).
    Figure Legend Snippet: Targeting USP7 activates local and systemic anti-tumor immunity. ( A ) Multicolor immunofluorescence detection of TAMs and CTLs in the TME of the P5091 group and the Control group. ( B-F ) Cytokines IFN-γ ( B ), TNF-α ( C ), IL-2 ( D ), IL-5 ( E ), and IL-6 ( F ) in the TME of each group were detected by Mul-Analyte Flow Assay Kit. ( G-K ) Proportion of Treg ( G ), CD4 + T ( H ), CD8 + T ( I ), Th1 ( J ), and CTLs ( K ) in the spleen of each group. Data are presented as the mean ± SEM (n = 7) for ( B-K ).

    Techniques Used: Immunofluorescence

    Targeting USP7 inhibits murine M2 phenotype and function in vitro . (A) The mRNA expression of common genes related to DUBs was detected by RT-PCR in M0, M1 (LPS/IFN-γ M1), and M2 (IL-4/13 M2 and IL-10 M2) induced from ANA-1. ( B ) The mRNA expression of USP7 in MΦs, M1, and M2 induced from BMDMs was detected by RT-PCR. ( C ) Western blotting showing the expression of USP7 in MΦ, M1, and M2 induced from BMDMs. ( D-E ) Flow cytometry analyses of the expression of CD206 in IL-4/13-induced BMDM M2 cells, which had been treated with P5091 (5 μM or 10 μM) for 24 h. Data are presented as the mean ± SEM (n = 3). ( F ) Detection of the expression of USP7 in BMDMs, which were transfected with either NC- or USP7-siRNA by western blotting. ( G ) The expression of CD206 in BMDMs of NC-siRNA and USP7-siRNA group was detected by flow cytometry. ( H-I ) Flow cytometry analyses of CFSE expression on the surface of CD8 + T cells in the presence of conditioned medium from either DMSO- or P5091- (5 μM or 10 μM) treated IL-4/13-induced BMDM M2 cells. Data are presented as the mean ± SEM (n = 3).
    Figure Legend Snippet: Targeting USP7 inhibits murine M2 phenotype and function in vitro . (A) The mRNA expression of common genes related to DUBs was detected by RT-PCR in M0, M1 (LPS/IFN-γ M1), and M2 (IL-4/13 M2 and IL-10 M2) induced from ANA-1. ( B ) The mRNA expression of USP7 in MΦs, M1, and M2 induced from BMDMs was detected by RT-PCR. ( C ) Western blotting showing the expression of USP7 in MΦ, M1, and M2 induced from BMDMs. ( D-E ) Flow cytometry analyses of the expression of CD206 in IL-4/13-induced BMDM M2 cells, which had been treated with P5091 (5 μM or 10 μM) for 24 h. Data are presented as the mean ± SEM (n = 3). ( F ) Detection of the expression of USP7 in BMDMs, which were transfected with either NC- or USP7-siRNA by western blotting. ( G ) The expression of CD206 in BMDMs of NC-siRNA and USP7-siRNA group was detected by flow cytometry. ( H-I ) Flow cytometry analyses of CFSE expression on the surface of CD8 + T cells in the presence of conditioned medium from either DMSO- or P5091- (5 μM or 10 μM) treated IL-4/13-induced BMDM M2 cells. Data are presented as the mean ± SEM (n = 3).

    Techniques Used: In Vitro, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Flow Cytometry, Transfection

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Boster Bio anti cd8 ab
    Phenotypic characteristics of grafts. Xenografts and allografts were recovered at POD 6. Relative mRNA expression of IL-2, IFN-γ, IL-4, and TNF-α in (A) xenograft and (B) allografts measured by qRT-PCR ( n = 3 mice/group). (C) Immunohistochemistry staining of CD20 + (brown) in xenograft and allograft; bar indicates 20 µm and quantified. (D) IgM in grafts were examined by immunohistochemistry staining; bar indicates 20 µm and quantified. Immunohistochemistry staining of CD4 + (brown), and <t>CD8</t> + (brown) in (E) xenograft and (H) allografts. (Original magnification: ×400). Data are presented as mean ± SEM from three independent experiments. * P
    Anti Cd8 Ab, supplied by Boster Bio, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti cd8 ab/product/Boster Bio
    Average 94 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    anti cd8 ab - by Bioz Stars, 2022-10
    94/100 stars
      Buy from Supplier

    92
    Boster Bio cd8 antibody
    Targeting USP7 can activate the p38 MAPK pathway to reprogram TAMs. ( A ) The strategy for sorting TAMs in TME by flow cytometry. ( B ) Heat maps illustrating the differentially expressed M1- and M2-related genes in TAMs between the P5091 group and the Control group based on the results of RNA sequencing. ( C ) RT-PCR further verifying the differentially expressed genes of sorted TAMs in each group. Data are presented as the mean ± SEM (n = 3). ( D ) KEGG analysis identifying 20 most obviously enriched pathways based on the differentially expressed genes of the two groups. ( E ) Western blotting detection of the expression of JNK, p-JNK, ERK1/2, p-ERK1/2, p38, p-p38, and β-actin in IL-4/13-BMDM M2 cells treated with P5091 (10 µM) at the indicated time points. ( F ) Flow cytometry analysis of CFSE expression on the surface of <t>CD8</t> + T cells in the presence of conditioned medium of IL-4/13-induced BMDM M2 cells from various indicated treatments. Treatments indicated: DMSO stimulation, P5091 (10 µM) stimulation, P5091 (10 µM) stimulation in the presence of inhibitors of p38 (SB203580, 10 µM), JNK (SP600125, 10 µM), Erk1/2 (U0126-EtOH, 10 µM). Data are presented as the mean ± SEM (n = 4).
    Cd8 Antibody, supplied by Boster Bio, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd8 antibody/product/Boster Bio
    Average 92 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd8 antibody - by Bioz Stars, 2022-10
    92/100 stars
      Buy from Supplier

    80
    Boster Bio cd8
    Validation of specific proteins during liver tumor development (Western blotting assay). These data showed significant upregulation of CD4 and <t>CD8</t> expressions in liver tumors when compared to those in nontumor liver cells. Notes: cancer vs normal, a P
    Cd8, supplied by Boster Bio, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cd8/product/Boster Bio
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cd8 - by Bioz Stars, 2022-10
    80/100 stars
      Buy from Supplier

    Image Search Results


    Phenotypic characteristics of grafts. Xenografts and allografts were recovered at POD 6. Relative mRNA expression of IL-2, IFN-γ, IL-4, and TNF-α in (A) xenograft and (B) allografts measured by qRT-PCR ( n = 3 mice/group). (C) Immunohistochemistry staining of CD20 + (brown) in xenograft and allograft; bar indicates 20 µm and quantified. (D) IgM in grafts were examined by immunohistochemistry staining; bar indicates 20 µm and quantified. Immunohistochemistry staining of CD4 + (brown), and CD8 + (brown) in (E) xenograft and (H) allografts. (Original magnification: ×400). Data are presented as mean ± SEM from three independent experiments. * P

    Journal: Cell Transplantation

    Article Title: Leflunomide Inhibits rat-to-Mouse Cardiac Xenograft Rejection by Suppressing Adaptive Immune Cell Response and NF-κB Signaling Activation

    doi: 10.1177/09636897211054503

    Figure Lengend Snippet: Phenotypic characteristics of grafts. Xenografts and allografts were recovered at POD 6. Relative mRNA expression of IL-2, IFN-γ, IL-4, and TNF-α in (A) xenograft and (B) allografts measured by qRT-PCR ( n = 3 mice/group). (C) Immunohistochemistry staining of CD20 + (brown) in xenograft and allograft; bar indicates 20 µm and quantified. (D) IgM in grafts were examined by immunohistochemistry staining; bar indicates 20 µm and quantified. Immunohistochemistry staining of CD4 + (brown), and CD8 + (brown) in (E) xenograft and (H) allografts. (Original magnification: ×400). Data are presented as mean ± SEM from three independent experiments. * P

    Article Snippet: Immunohistochemical staining was performed as described previously , the sections were then stained with primary antibody anti-CD4 Ab (Boster, Wuhan, China), anti-CD8 Ab (Boster), anti-CD20 Ab (Boster), anti-IgM (Boster), and secondary antibody goat anti-rabbit IgG (GB23303; Boster, Wuhan, China).

    Techniques: Expressing, Quantitative RT-PCR, Mouse Assay, Immunohistochemistry, Staining

    The effect of LEF treatment on CMR- and AVR- mediated immune responses. (A) MLR responses. Recipient splenocytes were isolated at POD 6 (responders) and irradiated naïve SD or BALB/c splenocytes (stimulators) were co-cultured for three days. Data are representative of three independent experiments. (B) Absolute numbers of splenocytes in LEF -treated and normal-saline treated mice recipients. Naïve C57BL/6 mice are shown for comparison ( n = 3 mice/group). (C) Representative proportion of CD4 + and CD8 + T cells in recipient splenocytes. A total of 1 × 10 6 splenocytes were isolated at POD 6, and the percentage of CD4 + and CD8 + T cells was determined by flow cytometry ( n = 3 mice/group). (D) Representative proportion of CD19 + B cells in recipient splenocytes. A total of 1 × 10 6 splenocytes were isolated at POD 6, and the percentage of CD19 + B cells was determined by flow cytometry ( n = 3 mice/group). (E) The number of CD3 + T cells in recipient spleen were determined by flow cytometry ( n = 3 mice/group). (F) The number of CD19 + B cells in recipient spleen were determined by flow cytometry ( n = 3 mice/group). (G) Representative proportion of donor-specific antibodies in recipient serum. Serum was collected from xenograft and allograft recipients at POD 6, and the percent of IgG1, IgG2a, and IgM was determined by flow cytometry ( n = 3 mice/group). (H) Serum levels of proinflammatory cytokines. Peripheral blood was collected at POD 6 and IFN-γ serum levels were measured by ELISA ( n = 3 mice/group). (I) CD3 + T cells were isolated from naïve C57BL/6 mice, and co-cultured with anti-CD3 and anti-CD28 monoclonal antibody in the absence and presence of LEF for 3 days, the supernatant was collected and measured by ELISA ( n = 3 separate experiments). Data are presented as the mean ± SEM of three independent experiments. * P

    Journal: Cell Transplantation

    Article Title: Leflunomide Inhibits rat-to-Mouse Cardiac Xenograft Rejection by Suppressing Adaptive Immune Cell Response and NF-κB Signaling Activation

    doi: 10.1177/09636897211054503

    Figure Lengend Snippet: The effect of LEF treatment on CMR- and AVR- mediated immune responses. (A) MLR responses. Recipient splenocytes were isolated at POD 6 (responders) and irradiated naïve SD or BALB/c splenocytes (stimulators) were co-cultured for three days. Data are representative of three independent experiments. (B) Absolute numbers of splenocytes in LEF -treated and normal-saline treated mice recipients. Naïve C57BL/6 mice are shown for comparison ( n = 3 mice/group). (C) Representative proportion of CD4 + and CD8 + T cells in recipient splenocytes. A total of 1 × 10 6 splenocytes were isolated at POD 6, and the percentage of CD4 + and CD8 + T cells was determined by flow cytometry ( n = 3 mice/group). (D) Representative proportion of CD19 + B cells in recipient splenocytes. A total of 1 × 10 6 splenocytes were isolated at POD 6, and the percentage of CD19 + B cells was determined by flow cytometry ( n = 3 mice/group). (E) The number of CD3 + T cells in recipient spleen were determined by flow cytometry ( n = 3 mice/group). (F) The number of CD19 + B cells in recipient spleen were determined by flow cytometry ( n = 3 mice/group). (G) Representative proportion of donor-specific antibodies in recipient serum. Serum was collected from xenograft and allograft recipients at POD 6, and the percent of IgG1, IgG2a, and IgM was determined by flow cytometry ( n = 3 mice/group). (H) Serum levels of proinflammatory cytokines. Peripheral blood was collected at POD 6 and IFN-γ serum levels were measured by ELISA ( n = 3 mice/group). (I) CD3 + T cells were isolated from naïve C57BL/6 mice, and co-cultured with anti-CD3 and anti-CD28 monoclonal antibody in the absence and presence of LEF for 3 days, the supernatant was collected and measured by ELISA ( n = 3 separate experiments). Data are presented as the mean ± SEM of three independent experiments. * P

    Article Snippet: Immunohistochemical staining was performed as described previously , the sections were then stained with primary antibody anti-CD4 Ab (Boster, Wuhan, China), anti-CD8 Ab (Boster), anti-CD20 Ab (Boster), anti-IgM (Boster), and secondary antibody goat anti-rabbit IgG (GB23303; Boster, Wuhan, China).

    Techniques: Isolation, Irradiation, Cell Culture, Mouse Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay

    Detection of mitochondrial damage and apoptosis in CD8 + T cells. Activated CD8 + T cells, co-cultured with Huh-7 cells for 3 d, were detected by flow cytometry. A: JC-1; B: Reactive oxygen species; C: Apoptosis. The data represent the mean ± SE; b P

    Journal: World Journal of Gastrointestinal Oncology

    Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

    doi: 10.4251/wjgo.v14.i6.1124

    Figure Lengend Snippet: Detection of mitochondrial damage and apoptosis in CD8 + T cells. Activated CD8 + T cells, co-cultured with Huh-7 cells for 3 d, were detected by flow cytometry. A: JC-1; B: Reactive oxygen species; C: Apoptosis. The data represent the mean ± SE; b P

    Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

    Techniques: Cell Culture, Flow Cytometry

    Regulation of CD8 + T cell function by carbonyl cyanide-3-chlorophenylhydrazone. A-C: Activated CD8 + T cells were cultured in RPMI 1640 supplemented with 5 μmol/L or 10 μmol/L carbonyl cyanide-3-chlorophenylhydrazone for 3 d, respectively. The levels of indicated molecules in CD8 + T cells were detected by flow cytometry. A: PD-1; B: PRF1; C: Granzyme B. The data represent the mean ± SE; d P

    Journal: World Journal of Gastrointestinal Oncology

    Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

    doi: 10.4251/wjgo.v14.i6.1124

    Figure Lengend Snippet: Regulation of CD8 + T cell function by carbonyl cyanide-3-chlorophenylhydrazone. A-C: Activated CD8 + T cells were cultured in RPMI 1640 supplemented with 5 μmol/L or 10 μmol/L carbonyl cyanide-3-chlorophenylhydrazone for 3 d, respectively. The levels of indicated molecules in CD8 + T cells were detected by flow cytometry. A: PD-1; B: PRF1; C: Granzyme B. The data represent the mean ± SE; d P

    Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

    Techniques: Cell Function Assay, Cell Culture, Flow Cytometry

    Regulation of CD8 + T cell function by Mdivi-1. A-C: Activated CD8 + T cells were co-cultured with Huh-7 cells in RPMI 1640 containing 5 μmol/L Mdivi-1 for 3 d. The levels of indicated molecules in CD8 + T cells were detected by flow cytometry. A: PD-1; B: PRF1; C: Granzyme B. The data represent the mean ± SE; c P

    Journal: World Journal of Gastrointestinal Oncology

    Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

    doi: 10.4251/wjgo.v14.i6.1124

    Figure Lengend Snippet: Regulation of CD8 + T cell function by Mdivi-1. A-C: Activated CD8 + T cells were co-cultured with Huh-7 cells in RPMI 1640 containing 5 μmol/L Mdivi-1 for 3 d. The levels of indicated molecules in CD8 + T cells were detected by flow cytometry. A: PD-1; B: PRF1; C: Granzyme B. The data represent the mean ± SE; c P

    Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

    Techniques: Cell Function Assay, Cell Culture, Flow Cytometry

    Effect of glutamine deprivation on CD8 + T cells co-cultured with hepatoma cells. Activated CD8 + T cells were co-cultured with Huh-7 cells in RPMI 1640 containing no glutamine (0Gln) for 3 d. The levels of indicated molecules in CD8 + T cells were detected by flow cytometry. A: PD-1; B: PRF1; C: Granzyme B; D: JC-1; E: Reactive oxygen species; F: Apoptosis. The data represent the mean ± SE; a P

    Journal: World Journal of Gastrointestinal Oncology

    Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

    doi: 10.4251/wjgo.v14.i6.1124

    Figure Lengend Snippet: Effect of glutamine deprivation on CD8 + T cells co-cultured with hepatoma cells. Activated CD8 + T cells were co-cultured with Huh-7 cells in RPMI 1640 containing no glutamine (0Gln) for 3 d. The levels of indicated molecules in CD8 + T cells were detected by flow cytometry. A: PD-1; B: PRF1; C: Granzyme B; D: JC-1; E: Reactive oxygen species; F: Apoptosis. The data represent the mean ± SE; a P

    Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

    Techniques: Cell Culture, Flow Cytometry

    PD-1 expression by infiltrating CD8 + T cells in liver tissues. Tissues were subjected to fluorescent double staining with anti-CD8 and anti-PD-1 antibodies. A: Hepatocellular carcinoma tissues; B: Paracancerous tissues. Scale bar: 400 μm.

    Journal: World Journal of Gastrointestinal Oncology

    Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

    doi: 10.4251/wjgo.v14.i6.1124

    Figure Lengend Snippet: PD-1 expression by infiltrating CD8 + T cells in liver tissues. Tissues were subjected to fluorescent double staining with anti-CD8 and anti-PD-1 antibodies. A: Hepatocellular carcinoma tissues; B: Paracancerous tissues. Scale bar: 400 μm.

    Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

    Techniques: Expressing, Double Staining

    Evaluations of CD8 + T cell function. Activated CD8 + T cells were co-cultured with Huh-7 cells for 3 d. Flow cytometry was used to detect the expression of indicated molecules in CD8 + T cells. A: PD-1; B: PRF1; C: Granzyme B (GZMB). D and E: Quantitative real-time polymerase chain reaction was used to detect the mRNA levels in CD8 + T cells (D: PRF1 ; E: GZMB ). The data represent the mean ± SE; a P

    Journal: World Journal of Gastrointestinal Oncology

    Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

    doi: 10.4251/wjgo.v14.i6.1124

    Figure Lengend Snippet: Evaluations of CD8 + T cell function. Activated CD8 + T cells were co-cultured with Huh-7 cells for 3 d. Flow cytometry was used to detect the expression of indicated molecules in CD8 + T cells. A: PD-1; B: PRF1; C: Granzyme B (GZMB). D and E: Quantitative real-time polymerase chain reaction was used to detect the mRNA levels in CD8 + T cells (D: PRF1 ; E: GZMB ). The data represent the mean ± SE; a P

    Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

    Techniques: Cell Function Assay, Cell Culture, Flow Cytometry, Expressing, Real-time Polymerase Chain Reaction

    Differential gene expression in CD8 + T cells. Activated CD8 + T cells were co-cultured with Huh-7 cells for 3 d. Total RNA was extracted from CD8 + T cells for transcriptome sequencing. A: GO enrichment plot of upregulated genes; B: GO enrichment plot of downregulated genes.

    Journal: World Journal of Gastrointestinal Oncology

    Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

    doi: 10.4251/wjgo.v14.i6.1124

    Figure Lengend Snippet: Differential gene expression in CD8 + T cells. Activated CD8 + T cells were co-cultured with Huh-7 cells for 3 d. Total RNA was extracted from CD8 + T cells for transcriptome sequencing. A: GO enrichment plot of upregulated genes; B: GO enrichment plot of downregulated genes.

    Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

    Techniques: Expressing, Cell Culture, Sequencing

    Transmission electron microscopy images of mitochondria in CD8 + T cells. Activated CD8 + T cells were co-cultured with Huh-7 cells for 3 d. The morphology of mitochondria in CD8 + T cells was detected by transmission electron microscopy. A: Control; B: Co-culture. Scale bar: 2 μm.

    Journal: World Journal of Gastrointestinal Oncology

    Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

    doi: 10.4251/wjgo.v14.i6.1124

    Figure Lengend Snippet: Transmission electron microscopy images of mitochondria in CD8 + T cells. Activated CD8 + T cells were co-cultured with Huh-7 cells for 3 d. The morphology of mitochondria in CD8 + T cells was detected by transmission electron microscopy. A: Control; B: Co-culture. Scale bar: 2 μm.

    Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

    Techniques: Transmission Assay, Electron Microscopy, Cell Culture, Co-Culture Assay

    CD8 + T cell infiltration in liver tissues. A: Hepatocellular carcinoma tissues; B: Paracancerous tissues. Scale bar: 200 μm.

    Journal: World Journal of Gastrointestinal Oncology

    Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

    doi: 10.4251/wjgo.v14.i6.1124

    Figure Lengend Snippet: CD8 + T cell infiltration in liver tissues. A: Hepatocellular carcinoma tissues; B: Paracancerous tissues. Scale bar: 200 μm.

    Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

    Techniques:

    Altered CD8 + T cell function by glutamine deficiency. Activated CD8 + T cells were cultured in RPMI 1640 containing no glutamine (0Gln) for 3 d. The levels of indicated molecules in CD8 + T cells were detected by flow cytometry. A: PD-1; B: PRF1; C: Granzyme B; D: JC-1; E: Reactive oxygen species; F: Apoptosis. The data represent the mean ± SE; b P

    Journal: World Journal of Gastrointestinal Oncology

    Article Title: Glutamine deprivation impairs function of infiltrating CD8+ T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

    doi: 10.4251/wjgo.v14.i6.1124

    Figure Lengend Snippet: Altered CD8 + T cell function by glutamine deficiency. Activated CD8 + T cells were cultured in RPMI 1640 containing no glutamine (0Gln) for 3 d. The levels of indicated molecules in CD8 + T cells were detected by flow cytometry. A: PD-1; B: PRF1; C: Granzyme B; D: JC-1; E: Reactive oxygen species; F: Apoptosis. The data represent the mean ± SE; b P

    Article Snippet: Immunofluorescence staining of paraffin-embedded human liver tissue sections was performed using anti-PD-1/CD279 (66220-1-Ig, Proteintech, Rosemont, IL, United States) and anti-CD8α (PB9249, Boster Bio, Pleasanton, CA, United States) antibodies.

    Techniques: Cell Function Assay, Cell Culture, Flow Cytometry

    Targeting USP7 can activate the p38 MAPK pathway to reprogram TAMs. ( A ) The strategy for sorting TAMs in TME by flow cytometry. ( B ) Heat maps illustrating the differentially expressed M1- and M2-related genes in TAMs between the P5091 group and the Control group based on the results of RNA sequencing. ( C ) RT-PCR further verifying the differentially expressed genes of sorted TAMs in each group. Data are presented as the mean ± SEM (n = 3). ( D ) KEGG analysis identifying 20 most obviously enriched pathways based on the differentially expressed genes of the two groups. ( E ) Western blotting detection of the expression of JNK, p-JNK, ERK1/2, p-ERK1/2, p38, p-p38, and β-actin in IL-4/13-BMDM M2 cells treated with P5091 (10 µM) at the indicated time points. ( F ) Flow cytometry analysis of CFSE expression on the surface of CD8 + T cells in the presence of conditioned medium of IL-4/13-induced BMDM M2 cells from various indicated treatments. Treatments indicated: DMSO stimulation, P5091 (10 µM) stimulation, P5091 (10 µM) stimulation in the presence of inhibitors of p38 (SB203580, 10 µM), JNK (SP600125, 10 µM), Erk1/2 (U0126-EtOH, 10 µM). Data are presented as the mean ± SEM (n = 4).

    Journal: Theranostics

    Article Title: USP7 targeting modulates anti-tumor immune response by reprogramming Tumor-associated Macrophages in Lung Cancer

    doi: 10.7150/thno.47137

    Figure Lengend Snippet: Targeting USP7 can activate the p38 MAPK pathway to reprogram TAMs. ( A ) The strategy for sorting TAMs in TME by flow cytometry. ( B ) Heat maps illustrating the differentially expressed M1- and M2-related genes in TAMs between the P5091 group and the Control group based on the results of RNA sequencing. ( C ) RT-PCR further verifying the differentially expressed genes of sorted TAMs in each group. Data are presented as the mean ± SEM (n = 3). ( D ) KEGG analysis identifying 20 most obviously enriched pathways based on the differentially expressed genes of the two groups. ( E ) Western blotting detection of the expression of JNK, p-JNK, ERK1/2, p-ERK1/2, p38, p-p38, and β-actin in IL-4/13-BMDM M2 cells treated with P5091 (10 µM) at the indicated time points. ( F ) Flow cytometry analysis of CFSE expression on the surface of CD8 + T cells in the presence of conditioned medium of IL-4/13-induced BMDM M2 cells from various indicated treatments. Treatments indicated: DMSO stimulation, P5091 (10 µM) stimulation, P5091 (10 µM) stimulation in the presence of inhibitors of p38 (SB203580, 10 µM), JNK (SP600125, 10 µM), Erk1/2 (U0126-EtOH, 10 µM). Data are presented as the mean ± SEM (n = 4).

    Article Snippet: CD8 antibody was purchased from BOSTER Biological Technology Co., Ltd (Wuhan, China).

    Techniques: Flow Cytometry, RNA Sequencing Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing

    Targeting USP7 inhibits tumor growth and induces local anti-tumor immunity in vivo . ( A ) Harvested and photographed tumors in the P5091 and the Control group. ( B ) Lewis tumor growth following P5091 or Vehicle treatment in vivo. Data are presented as the mean ± SEM (n = 6 per group). ( C ) Spider diagram of the tumor volume growth in each mouse from the P5091 group and the Control group. ( D ) Gating strategy for detection of the TAMs by flow cytometry. ( E-G ) Proportions of M1 ( E ) and M2 ( F ), and M1/M2 ratio ( G ) in the TME of the P5091 group and the Control group. ( H-M ) Percentages of MDSC ( H ), Treg ( I ), CD4 + T ( J ), CD8 + T ( K ), Th1 ( L ), and CTLs ( M ) within the TME in each group. Data are presented as the mean ± SEM (n = 7) for ( E-M ).

    Journal: Theranostics

    Article Title: USP7 targeting modulates anti-tumor immune response by reprogramming Tumor-associated Macrophages in Lung Cancer

    doi: 10.7150/thno.47137

    Figure Lengend Snippet: Targeting USP7 inhibits tumor growth and induces local anti-tumor immunity in vivo . ( A ) Harvested and photographed tumors in the P5091 and the Control group. ( B ) Lewis tumor growth following P5091 or Vehicle treatment in vivo. Data are presented as the mean ± SEM (n = 6 per group). ( C ) Spider diagram of the tumor volume growth in each mouse from the P5091 group and the Control group. ( D ) Gating strategy for detection of the TAMs by flow cytometry. ( E-G ) Proportions of M1 ( E ) and M2 ( F ), and M1/M2 ratio ( G ) in the TME of the P5091 group and the Control group. ( H-M ) Percentages of MDSC ( H ), Treg ( I ), CD4 + T ( J ), CD8 + T ( K ), Th1 ( L ), and CTLs ( M ) within the TME in each group. Data are presented as the mean ± SEM (n = 7) for ( E-M ).

    Article Snippet: CD8 antibody was purchased from BOSTER Biological Technology Co., Ltd (Wuhan, China).

    Techniques: In Vivo, Flow Cytometry

    Targeting USP7 activates local and systemic anti-tumor immunity. ( A ) Multicolor immunofluorescence detection of TAMs and CTLs in the TME of the P5091 group and the Control group. ( B-F ) Cytokines IFN-γ ( B ), TNF-α ( C ), IL-2 ( D ), IL-5 ( E ), and IL-6 ( F ) in the TME of each group were detected by Mul-Analyte Flow Assay Kit. ( G-K ) Proportion of Treg ( G ), CD4 + T ( H ), CD8 + T ( I ), Th1 ( J ), and CTLs ( K ) in the spleen of each group. Data are presented as the mean ± SEM (n = 7) for ( B-K ).

    Journal: Theranostics

    Article Title: USP7 targeting modulates anti-tumor immune response by reprogramming Tumor-associated Macrophages in Lung Cancer

    doi: 10.7150/thno.47137

    Figure Lengend Snippet: Targeting USP7 activates local and systemic anti-tumor immunity. ( A ) Multicolor immunofluorescence detection of TAMs and CTLs in the TME of the P5091 group and the Control group. ( B-F ) Cytokines IFN-γ ( B ), TNF-α ( C ), IL-2 ( D ), IL-5 ( E ), and IL-6 ( F ) in the TME of each group were detected by Mul-Analyte Flow Assay Kit. ( G-K ) Proportion of Treg ( G ), CD4 + T ( H ), CD8 + T ( I ), Th1 ( J ), and CTLs ( K ) in the spleen of each group. Data are presented as the mean ± SEM (n = 7) for ( B-K ).

    Article Snippet: CD8 antibody was purchased from BOSTER Biological Technology Co., Ltd (Wuhan, China).

    Techniques: Immunofluorescence

    Targeting USP7 inhibits murine M2 phenotype and function in vitro . (A) The mRNA expression of common genes related to DUBs was detected by RT-PCR in M0, M1 (LPS/IFN-γ M1), and M2 (IL-4/13 M2 and IL-10 M2) induced from ANA-1. ( B ) The mRNA expression of USP7 in MΦs, M1, and M2 induced from BMDMs was detected by RT-PCR. ( C ) Western blotting showing the expression of USP7 in MΦ, M1, and M2 induced from BMDMs. ( D-E ) Flow cytometry analyses of the expression of CD206 in IL-4/13-induced BMDM M2 cells, which had been treated with P5091 (5 μM or 10 μM) for 24 h. Data are presented as the mean ± SEM (n = 3). ( F ) Detection of the expression of USP7 in BMDMs, which were transfected with either NC- or USP7-siRNA by western blotting. ( G ) The expression of CD206 in BMDMs of NC-siRNA and USP7-siRNA group was detected by flow cytometry. ( H-I ) Flow cytometry analyses of CFSE expression on the surface of CD8 + T cells in the presence of conditioned medium from either DMSO- or P5091- (5 μM or 10 μM) treated IL-4/13-induced BMDM M2 cells. Data are presented as the mean ± SEM (n = 3).

    Journal: Theranostics

    Article Title: USP7 targeting modulates anti-tumor immune response by reprogramming Tumor-associated Macrophages in Lung Cancer

    doi: 10.7150/thno.47137

    Figure Lengend Snippet: Targeting USP7 inhibits murine M2 phenotype and function in vitro . (A) The mRNA expression of common genes related to DUBs was detected by RT-PCR in M0, M1 (LPS/IFN-γ M1), and M2 (IL-4/13 M2 and IL-10 M2) induced from ANA-1. ( B ) The mRNA expression of USP7 in MΦs, M1, and M2 induced from BMDMs was detected by RT-PCR. ( C ) Western blotting showing the expression of USP7 in MΦ, M1, and M2 induced from BMDMs. ( D-E ) Flow cytometry analyses of the expression of CD206 in IL-4/13-induced BMDM M2 cells, which had been treated with P5091 (5 μM or 10 μM) for 24 h. Data are presented as the mean ± SEM (n = 3). ( F ) Detection of the expression of USP7 in BMDMs, which were transfected with either NC- or USP7-siRNA by western blotting. ( G ) The expression of CD206 in BMDMs of NC-siRNA and USP7-siRNA group was detected by flow cytometry. ( H-I ) Flow cytometry analyses of CFSE expression on the surface of CD8 + T cells in the presence of conditioned medium from either DMSO- or P5091- (5 μM or 10 μM) treated IL-4/13-induced BMDM M2 cells. Data are presented as the mean ± SEM (n = 3).

    Article Snippet: CD8 antibody was purchased from BOSTER Biological Technology Co., Ltd (Wuhan, China).

    Techniques: In Vitro, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Flow Cytometry, Transfection

    Validation of specific proteins during liver tumor development (Western blotting assay). These data showed significant upregulation of CD4 and CD8 expressions in liver tumors when compared to those in nontumor liver cells. Notes: cancer vs normal, a P

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: MicroRNA-421 mediates immunosensitivity in late-stage human liver cancer

    doi:

    Figure Lengend Snippet: Validation of specific proteins during liver tumor development (Western blotting assay). These data showed significant upregulation of CD4 and CD8 expressions in liver tumors when compared to those in nontumor liver cells. Notes: cancer vs normal, a P

    Article Snippet: For immunoblotting analysis, equal amount of protein (40 µg/lane in each nontumor or tumor sample) was subjected to electrophoretic separation with SDS-PAGE, followed by targeting protein being blotted to membrane and incubated with antibodies against CD4 and CD8 (1:200, Boster, Wuhan, China).

    Techniques: Western Blot