Journal: Cell reports

Article Title: The aryl hydrocarbon receptor cell intrinsically promotes resident memory CD8 + T cell differentiation and function

doi: 10.1016/j.celrep.2022.111963

Figure Lengend Snippet: (A and B) Flow cytometry analysis of Ahr expression (GFP) in CD8 + T cells of Ahr +/+ and Ahr dCAIR/+ mice was performed. Histogram plot of GFP( Ahr ) in CD8 + T cells isolated from spleen (Sp), peripheral lymph node (pLN), mesenteric lymph node (mLN), small intestine lamina propria (LPL), or intraepithelial lymphocytes (IEL) (A). Quantification of ΔgMFI in Ahr dCAIR/+ compared with GFP-negative Ahr +/+ mice (B). (C and D) Flow cytometry quantification of ΔgMFI of Ahr -GFP in different CD8 + T cell populations isolated from LPL including CD44 − CD62L + (T naive ), CD127 − KLRG1 − (T EE ), CD127 − KLRG1 + (T SLE ), CD127 + KLRG1 − (T MPE ) (C), and CD44 + CD62L + (T CM ), CD44 + CD62L − (T EM ), and CD69 + CD103 + (T RM ) (D). Data are shown as mean ± SEM (n = 3 mice per group). Data are representative of two independent experiments. See also .

Article Snippet: Anti-human CD8 – APC-Cyanine7 (clone SK1) , TONBO biosciences , Cat# 25-0087-T025; RRID: AB_2848136.

Techniques: Flow Cytometry, Expressing, Isolation

Journal: Cell reports

Article Title: The aryl hydrocarbon receptor cell intrinsically promotes resident memory CD8 + T cell differentiation and function

doi: 10.1016/j.celrep.2022.111963

Figure Lengend Snippet: (A–E and H) RNA-seq analysis of IEL resident CD8 + T cells isolated from Ahr f/f and Ahr f/f Cd8 cre mice and statistics (q values shown) were calculated via DESeq2 differential expression analysis, and FPKM values were quantified using RSEM. MA plot highlighting genes upregulated (red) and downregulated (blue) in Ahr f/f Cd8 cre compared with Ahr f/f (A). Gene set enrichment analysis of resident and circulating core gene signatures (B). Heatmap depicting (fold change >1.5) genes enriched in respective signatures (C). FPKM expression values for transcriptional regulators (D) and secreted factors associated with cell function (E). (F and G) Flow cytometry quantification of granzyme B protein levels of IEL resident CD8 + T cells isolated from Ahr f/f and Ahr f/f Cd8 cre mice. The percentages (F) and total cell number (n = 5 mice per group) (G) are shown. Data are representative of two independent experiments. (H) FPKM expression values for proliferation, cell cycle, and apoptosis genes. (I–K) Naive CD8 + T cells were isolated from Ahr +/+ and Ahr − / − mice and then subjected to in vitro T RM -like differentiation culture conditions. RNA was isolated for qRT-PCR expression analysis of Ahrr (I). Flow cytometry quantification of CD69 + CD103 + in vitro T RM -like cells (J) and CD44 + CD62L + in vitro T CM -like population frequency (K). Data are shown as mean ± SEM (n = 3 mice per group). Data are representative of three independent experiments. See also , , and .

Article Snippet: Anti-human CD8 – APC-Cyanine7 (clone SK1) , TONBO biosciences , Cat# 25-0087-T025; RRID: AB_2848136.

Techniques: RNA Sequencing, Isolation, Quantitative Proteomics, Expressing, Cell Function Assay, Flow Cytometry, In Vitro, Quantitative RT-PCR

Journal: Cell reports

Article Title: The aryl hydrocarbon receptor cell intrinsically promotes resident memory CD8 + T cell differentiation and function

doi: 10.1016/j.celrep.2022.111963

Figure Lengend Snippet: (A and B) ChIP-seq of Ahr was performed using in vitro T RM -like CD8 + T cells. Analysis of Ahr binding was performed, and pie chart of peak annotation (A) as well as top seven enriched transcription factor motifs (B) are shown. (C and D) Binding and Expression Target Analysis (BETA) was performed to integrate RNA-seq and ChIP-seq data. Visualization of transcription factor activating/repressive function prediction (C) and rank product volcano plot depicting top direct target candidates (D). (E and F) Ahr − / − in vitro T RM -like CD8 + T cells transduced with retroviral constructs encoding MIG-EV (empty vector), MIG-Ahr, MIG-Y9A, or MIG-DbHLH. The cells were treated with DMSO or FICZ on day 3. On day 5, CD69 and CD103 expression was analyzed by flow cytometry (E), and RNA was isolated for qRT-PCR expression analysis of Ahr direct target gene Ahrr (F). Data are shown as mean ± SEM (n = 3 technical replicates per group). Data are representative of two independent experiments. (G) Flow cytometry analysis of CD69 and CD103 expression in Ahr − / − in vitro T RM -like CD8 + T cells transduced with retroviral constructs encoding MIG-EV, MIG-Ahr, hCD2-EV, or hCD2-Blimp1. Data are representative of two independent experiments. See also .

Article Snippet: Anti-human CD8 – APC-Cyanine7 (clone SK1) , TONBO biosciences , Cat# 25-0087-T025; RRID: AB_2848136.

Techniques: ChIP-sequencing, In Vitro, Binding Assay, Expressing, RNA Sequencing, Transduction, Retroviral, Construct, Plasmid Preparation, Flow Cytometry, Isolation, Quantitative RT-PCR

Journal: Cell reports

Article Title: The aryl hydrocarbon receptor cell intrinsically promotes resident memory CD8 + T cell differentiation and function

doi: 10.1016/j.celrep.2022.111963

Figure Lengend Snippet: (A–F) Analysis of antigen-specific (OTI) CD8 + T cell response in the IEL during oral L . m .-OVA infection was performed as depicted in the schematic (A). Flow cytometry quantification of the percentage of Ahr +/+ vs. Ahr − / − OTI cells on day 9, 20, and 34 post infection (B). Flow cytometry quantification of memory precursor populations based on expression of CD127 and KLRG1 (C) as well as CD69 and CD103 (D) in Ahr +/+ vs. Ahr − / − OTI cells present in the IEL on day 9 post infection. Flow cytometry analysis of CD45.1 and GzmB gated on OTI cells depicting CD45 . 1 + Ahr +/+ (CD45.1/.2) and CD45 . 1 − Ahr − / − (CD45.2/.2) OTI cells production of granzyme B on day 34 post infection (E). Quantification of granzyme B + OTI T cells in the mice of indicated genotypes (F). (G–J) L . m .-OVA re-infection was performed and Ahr +/+ vs. Ahr − / − OTI IEL resident CD8 + T cells analyzed on day 3 post re-infection. Flow cytometry analysis of CD45.1 and CD45.2 depicting percentage of Ahr +/+ (CD45.1/.2) and Ahr − / − (CD45.2/.2) OTI IEL resident CD8 + T cells as well as (G) quantification of percentage are shown (H). Flow cytometry analysis (I) and quantification (J) of granzyme B production in analyzed cells. Data are compiled from two independent experiments and shown as mean ± SEM (n = 3–6 replicates per group). See also .

Article Snippet: Anti-human CD8 – APC-Cyanine7 (clone SK1) , TONBO biosciences , Cat# 25-0087-T025; RRID: AB_2848136.

Techniques: Infection, Flow Cytometry, Expressing

Journal: Cell reports

Article Title: The aryl hydrocarbon receptor cell intrinsically promotes resident memory CD8 + T cell differentiation and function

doi: 10.1016/j.celrep.2022.111963

Figure Lengend Snippet: (A–D) scRNA-seq analysis of Ahr +/+ (n = 2) vs. Ahr − / − (n = 2) TIL CD8 + T cells was performed. UMAP dimensionality reduction and cluster visualization (left) as well as pie chart frequency depiction (right) color-coded to represent cluster ID (A). Pseudotime visualization (B) and quantification (C). Differential gene expression depicted via color intensity as average expression and circle size as percent expressed in CD8 + T cells (D). (E–H) Ahr f/f and Ahr f/f Cd8 cre mice were inoculated subcutaneously with B16F10 mouse melanoma and tumor size monitored (E). At endpoint, tumor weight was quantified (F), and TILs were isolated for flow cytometry analysis. Pie chart visualization depicting polyfunctionality of TIL CD8 + T cells in mice with indicated genotypes (G). Polyfunctionality score quantification (triple-positive plus double-positive minus triple-negative divided by total cells) of TIL CD8 + T cells isolated from Ahr f/f and Ahr f/f Cd8 cre tumor-bearing mice (H). Data are compiled from two independent experiments and are shown as mean ± SEM (n = 6 to 7 replicates per group). See also .

Article Snippet: Anti-human CD8 – APC-Cyanine7 (clone SK1) , TONBO biosciences , Cat# 25-0087-T025; RRID: AB_2848136.

Techniques: Gene Expression, Expressing, Isolation, Flow Cytometry

Journal: Cell reports

Article Title: The aryl hydrocarbon receptor cell intrinsically promotes resident memory CD8 + T cell differentiation and function

doi: 10.1016/j.celrep.2022.111963

Figure Lengend Snippet: (A–C) Human peripheral blood (PBMC) vs. IEL CD8 + T cells were analyzed via flow cytometry. Staining of CD45RA and CD45RO (top), CD103 and CD69 (bottom) (A), as well as T-BET and AHR (B) in tissue fractions as indicated in the figure. Quantification of AHR protein levels (gMFI) in human PBMC, LPL, and IEL CD8 + T cells (C). Data are compiled from three independent experiments and shown as mean ± SEM (n = 4 replicates per group). (D–G) Human naive CD8 + T cells were isolated from PBMCs and then subjected to in vitro T RM -like differentiation culture conditions. The cells were given differentiation cytokines and treated with DMSO (control), FICZ, or CH223191 on day 2. The assay was collected on day 4, and flow cytometry quantification of CD103 expression was performed (D). Data are compiled from three independent experiments and are shown as mean ± SEM (n = 4 replicates per group). RNA was isolated for qRT-PCR expression analysis of AHR (E), AHR direct target gene AHRR (F), and GZMB (G). Data are shown as mean ± SEM (n = 3 technical replicates per group). Data are representative of three independent experiments.

Article Snippet: Anti-human CD8 – APC-Cyanine7 (clone SK1) , TONBO biosciences , Cat# 25-0087-T025; RRID: AB_2848136.

Techniques: Flow Cytometry, Staining, Isolation, In Vitro, Control, Expressing, Quantitative RT-PCR

Journal: Cell reports

Article Title: The aryl hydrocarbon receptor cell intrinsically promotes resident memory CD8 + T cell differentiation and function

doi: 10.1016/j.celrep.2022.111963

Figure Lengend Snippet:

Article Snippet: Anti-human CD8 – APC-Cyanine7 (clone SK1) , TONBO biosciences , Cat# 25-0087-T025; RRID: AB_2848136.

Techniques: Virus, Isolation, Recombinant, SYBR Green Assay, DNA Library Preparation, Reverse Transcription, Staining, Transgenic Assay, Plasmid Preparation, Software

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: A IOD of ACAT2 expression in CC tissues and adjacent tissues was examined using immunohistochemical staining ( n = 47 biologically independent samples). IOD of DHCR7 B and MSMO1 C expression in CC patients with high ( n = 27 biologically independent samples) or low ( n = 20 biologically independent samples) expression of ACAT2 was examined using immunohistochemical staining. The number of activated CD8 T cells (CD8A + GZMB + ) D or activated NK cells (CD56 + GZMB + ) E infiltrated in the tumor tissues of patients with high ( n = 27 biologically independent samples) and low ACAT2 ( n = 20 biologically independent samples) expression was detected. Data represent mean ± SEM. Statistical analysis was performed using the paired A or unpaired ( B – E ) t-test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: ACAT2 expression in HCeEpiC and CC cell lines was examined using RT-qPCR A and Western blot analysis B ( n = 5 independent experiments). C ACAT2, DHCR7, and MSMO1 expression in CC cells after infection with Scramble-sh, ACAT2-sh #1, and ACAT2-sh #2 was examined using Western blot analysis ( n = 5 independent experiments). D Detection of total cholesterol, free cholesterol, and cholesteryl ester levels in CC cells ( n = 5 independent experiments). The proliferation of CC cells was examined using CCK8 ( E ) and colony formation assays F (n = 5 independent experiments). G CC cells were co-cultured with (E: T = 3:1) with NK cells or CD8 T cells for 6 h, respectively, and the death of CC cells was detected ( n = 5 independent experiments). H IFN-γ and GZMB released from immune cells in a co-culture system with CC cells were examined using ELISA ( n = 5 independent experiments). Data represent mean ± SEM. Statistical analysis was performed using the one-way ( A , B ) or two-way ( C - H ) ANOVA, followed by Tukey’s multiple comparisons test ( A – H ).

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Infection, Cell Culture, Co-Culture Assay, Enzyme-linked Immunosorbent Assay

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: A ACAT2 knockdown efficiency in U14 cells was examined using western blot analysis ( n = 10 independent experiments). B Volume changes of transplanted tumors in mice subcutaneously inoculated with U14 cells (n = 10 animals). C The images and weight of the tumors harvested on day 21 ( n = 10 animals). D Protein expression of ACAT2, MSMO1, DHCR7, and PCNA in transplanted tumors was examined using western blot analysis ( n = 10 animals). E Detection of total cholesterol, free cholesterol, and cholesteryl ester levels in transplanted tumors ( n = 10 animals). The gating strategy for GZMB + NK cells and CD8 + T cells F and quantification G were analyzed using flow cytometry ( n = 10 animals). H Survival of mice over 60 days after subcutaneous inoculation of U14 cells was analyzed using the log-rank test ( n = 20 animals). Data represent mean ± SEM. Statistical analysis was performed using the one-way ( A , C , E , G ) or two-way ( B , D ) ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Knockdown, Western Blot, Expressing, Flow Cytometry

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: The proliferation of CC cells was examined using CCK8 A and colony formation assays B ( n = 5 independent experiments). C CC cells were co-cultured with (E: T = 3:1) with NK cells or CD8 + T cells, and the death of CC cells was detected ( n = 5 independent experiments). D IFN-γ and GZMB released from immune cells in a co-culture system with CC cells were examined using ELISA ( n = 5 independent experiments). E TGF-β1 released by CC cells was examined using ELISA ( n = 5 independent experiments). F PD-L1 expression levels in CC cells were observed using immunofluorescence staining ( n = 5 independent experiments). Data represent mean ± SEM. Statistical analysis was performed using the two-way ( A – F ) ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Cell Culture, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence, Staining

Journal: Communications Biology

Article Title: SREBF2 enhances lipid metabolism and represses anti-tumor immune responses in cervical cancer by increasing ACAT2

doi: 10.1038/s42003-026-09678-9

Figure Lengend Snippet: A Volume changes of transplanted tumors in mice subcutaneously inoculated with U14 cells ( n = 5 animals). B The images and weight of the tumors harvested on day 21 ( n = 5 animals). The gating strategy for GZMB + NK cells and CD8 + T cells C and quantification D were analyzed using flow cytometry ( n = 5 animals). Data represent mean ± SEM. Statistical analysis was performed using the one-way ( B , D ) or two-way A ANOVA, followed by Tukey’s multiple comparisons test.

Article Snippet: The cell suspension (100 μL) was incubated with BeyoFC Fc Receptor Blocking Solution (C1755, Beyotime) for 10 min at 4 °C and with primary antibodies, including FITC-coupled CD3 antibody (1:100, FITC-65077, ProteinTech, RRID: AB_2883763), PE-coupled NK1.1 antibody (1:100, PE-65138, ProteinTech, RRID: AB_2883920), and APC-coupled CD8A antibody (1:100, APC-65069, ProteinTech, RRID: AB_2882970) for 1 h at 4 °C.

Techniques: Flow Cytometry

Journal: bioRxiv

Article Title: Balancing activation and costimulation of CAR tunes signaling dynamics and enhances therapeutic potency

doi: 10.1101/2022.03.01.482445

Figure Lengend Snippet: CD19-1XX CAR-T cells show better anti-tumor activity in a pancreatic tumor model in vivo. (a) Timeline of of CD19 CAR-T treatment in mouse model. (b) Panc-1 CD19+ -bearing mice were treated with 1 × 10 6 CD19 CAR-T cells as indicated and tumor burden (tumor volume) of mice was measured at indicated days (n=4 mice each group). For mouse experiments, Control, PBS. (c) Tumor tissue from mice treated with indicated CAR-T cells. (d) Tumor tissue from mice treated with indicated CAR-T cells was sliced and stained with antibodies against human CD4 and CD8 (n=4). The percentage of CD4 + (e) and CD8 + (f) T cells in tumor tissue from different groups of mice (n=4). All data are mean ± SEM.

Article Snippet: Anti-Human CD4 antibody (BD, Horizon, BUV395), anti-Human CD8 antibody (BD, Pharmin, APC-CY™7), anti-Human CD279(PD-1) antibody (Invitrogen, eBioscience, PE-Cyanine7), anti-Human CD223(LAG-3) antibody (Invitrogen, eBioscience,, Percp-efluor™710), anti-Human CD45RA antibody (Invitrogen, eBioscience, FITC), anti-Human CD62L antibody (Invitrogen, eBioscience, efluor450).

Techniques: Activity Assay, In Vivo, Staining

Journal: bioRxiv

Article Title: Balancing activation and costimulation of CAR tunes signaling dynamics and enhances therapeutic potency

doi: 10.1101/2022.03.01.482445

Figure Lengend Snippet: 1XX modification of AXL CAR enhance its anti-tumor activity in a melanoma tumor model. (a) Timeline of of AXL CAR-T treatment in mouse model. (b) A375-bearing mice were treated with 2.5 × 10 6 AXL CAR-T cells and tumor burden (tumor volume) of mice were measured at indicated days (n=4, 5, 5 mice in respective groups). Control, PBS. (c) Tumor tissue of mice treated with indicated CAR-T cells (n=4, 5, 5). (d) The weight of tumors from different groups of mice (n=4, 5, 5). Number of CAR-T(e), CD4 + CAR-T cells (f) and CD8 + CAR-T cells (g) in spleen of mice 20 days post CAR-T infusion (n=5, 3). (h-k) Phenotype of CAR-T cells in the spleen of mice 20 days after CAR-T infusion as demonstrated by the percentage of T CM (CD45RA - CD62L + ) and T EFF (CD45RA + CD62L - ) cells (n=5, 3). All data are mean ± SEM.

Article Snippet: Anti-Human CD4 antibody (BD, Horizon, BUV395), anti-Human CD8 antibody (BD, Pharmin, APC-CY™7), anti-Human CD279(PD-1) antibody (Invitrogen, eBioscience, PE-Cyanine7), anti-Human CD223(LAG-3) antibody (Invitrogen, eBioscience,, Percp-efluor™710), anti-Human CD45RA antibody (Invitrogen, eBioscience, FITC), anti-Human CD62L antibody (Invitrogen, eBioscience, efluor450).

Techniques: Modification, Activity Assay

Journal: Canadian Urological Association Journal

Article Title: Clinicopathological significance of galectin-1 expression and percentage of galectin-1-expressing T cells in clear-cell renal cell carcinoma

doi: 10.5489/cuaj.4573

Figure Lengend Snippet: Proportions of galectin-1-expressing CD4+ lymphocytes and galectin-1-expressing CD8+ lymphocytes in peripheral blood of patients with renal cell carcinoma and patients with non-renal cell carcinoma. (A) Proportions of galectin-1-expressing CD4+ lymphocytes; (B) proportions of galectin-1-expressing CD8+ lymphocytes.

Article Snippet: Cells were stained with mononuclear antibody PE-labeled anti-human CD4 and mononuclear antibody (APC)-labeled anti-human CD8 (BD Pharmingen, San Diego, CA, U.S.) for 15 minutes in the dark at room temperature and then washed twice with PBS.

Techniques: Expressing

Journal: Canadian Urological Association Journal

Article Title: Clinicopathological significance of galectin-1 expression and percentage of galectin-1-expressing T cells in clear-cell renal cell carcinoma

doi: 10.5489/cuaj.4573

Figure Lengend Snippet: Representative flow cytometric pattern showing intracellular galectin-1 expression of peripheral blood CD4+ and CD8+ lymphocytes in patients with renal cell carcinoma (RCC) and oncocytoma. (A) RCC patient; (B) oncocytoma patient; (C) isotype control.

Article Snippet: Cells were stained with mononuclear antibody PE-labeled anti-human CD4 and mononuclear antibody (APC)-labeled anti-human CD8 (BD Pharmingen, San Diego, CA, U.S.) for 15 minutes in the dark at room temperature and then washed twice with PBS.

Techniques: Expressing