cd44 apc cy7  (Cytek Biosciences)

Journal: bioRxiv

Article Title: Immunogenicity and protective efficacy of an intranasal neuraminidase-based influenza virus vaccine adjuvanted with bacterial cell membrane-derived adjuvants

doi: 10.1101/2025.02.26.640278

Figure Lengend Snippet: a. Representative flow cytometry plots showing the percentage of IL-2- and TNFα-producing CD4 + EM T-lymphocytes in mouse lungs and spleens 10 days after the booster immunization (gated on live CD3 + CD19 ‒ CD4 + CD62L ‒ CD44 + cells). Cells were incubated with overlapping peptides, covering the whole sequence of N1 NA-protein for 6h in the presence of Brefeldin A and co-stimulatory anti-CD28 antibodies. b. Percentage of different cytokine-producing cell populations within the total CD4 + EM T-cell subset after background subtraction. Average and SE values are shown. Statistical analyses were performed using one-way ANOVA with Tukey posthoc test. P-values for the groups with statistically significant differences are shown on the plots.

Article Snippet: For the analysis of T- and B-cell populations in lungs, spleen, and NALTs, cells were stained with the antibody cocktail, including the following antibodies: CD38-AF488 (0.125 μl, BioLegend), PD-1-PE/Dazzle (0.5 μl, BioLegend), CD273-RB744 (0.25 μl, BD Biosciences), CD138-PE (0.25 μl, BioLegend), CD103-PE-Cy5.5 (0.5 μl, BioLegend), CD44-PE/Cy7 (0.25 μl, Tonbo Biosciences), GL7-APC (0.25 μl, BioLegend), CD69-AF700 (1 μl, BioLegend), CD62L-APC/Cy7 (0.125 μl, BioLegend), CD185-BV421 (0.5 μl, BioLegend), CD80-BV605 (1 μl, BioLegend), IgD-BV650 (0.25 μl, BioLegend), CD3-BV711 (0.5 μl, BioLegend), CD8-BV785 (0.25 μl, BioLegend), CD4-BUV496 (0.125 μl, BD Biosciences), IgM-BUV737 (1 μl, BD Biosciences), CD19-BUV805 (0.125 μl, BD Biosciences).

Techniques: Flow Cytometry, Incubation, Sequencing

Journal: bioRxiv

Article Title: Immunogenicity and protective efficacy of an intranasal neuraminidase-based influenza virus vaccine adjuvanted with bacterial cell membrane-derived adjuvants

doi: 10.1101/2025.02.26.640278

Figure Lengend Snippet: a. Body weight dynamics and survival after the heterologous challenge with 5 × LD 50 of A/bald eagle/Florida/W22-134-OP/2022 (H5N1 reassortant with A/Puerto Rico/8/1934 vaccine backbone) and A/New Caledonia/20/1999 (H1N1) (n = 8). b. Viral titers in lungs and nasal turbinates on day 4 after the challenge (n = 4). c. Representative flow cytometry plots showing the percentage of IL-2- and TNFα-producing CD4 + EM T-lymphocytes (gated on live CD3 + CD19 ‒ CD4 + CD62L ‒ CD44 + cells) in mouse lungs and spleens 5 days after the challenge with 0.1 × LD 50 of A/bald eagle/Florida/W22-134-OP/2022 (H5N1). Cells were incubated with overlapping peptides, covering the whole sequence of N1 NA-protein for 6h in the presence of Brefeldin A and co-stimulatory anti-CD28 antibodies. d. Percentage of different cytokine-producing cell populations within the total CD4 + EM T-cell subset after background subtraction. Average and SE values are shown. Statistical analyses were performed using one-way ANOVA with Tukey posthoc test. P-values for the groups with statistically significant differences are shown on the plots.

Article Snippet: For the analysis of T- and B-cell populations in lungs, spleen, and NALTs, cells were stained with the antibody cocktail, including the following antibodies: CD38-AF488 (0.125 μl, BioLegend), PD-1-PE/Dazzle (0.5 μl, BioLegend), CD273-RB744 (0.25 μl, BD Biosciences), CD138-PE (0.25 μl, BioLegend), CD103-PE-Cy5.5 (0.5 μl, BioLegend), CD44-PE/Cy7 (0.25 μl, Tonbo Biosciences), GL7-APC (0.25 μl, BioLegend), CD69-AF700 (1 μl, BioLegend), CD62L-APC/Cy7 (0.125 μl, BioLegend), CD185-BV421 (0.5 μl, BioLegend), CD80-BV605 (1 μl, BioLegend), IgD-BV650 (0.25 μl, BioLegend), CD3-BV711 (0.5 μl, BioLegend), CD8-BV785 (0.25 μl, BioLegend), CD4-BUV496 (0.125 μl, BD Biosciences), IgM-BUV737 (1 μl, BD Biosciences), CD19-BUV805 (0.125 μl, BD Biosciences).

Techniques: Flow Cytometry, Incubation, Sequencing

Journal: eBioMedicine

Article Title: Intranasal recombinant protein subunit vaccine targeting TLR3 induces respiratory tract IgA and CD8 T cell responses and protects against respiratory virus infection

doi: 10.1016/j.ebiom.2025.105615

Figure Lengend Snippet: Intranasal administration of CAF09b adjuvanted spike protein elicits IgA responses and antigen-specific CD8 T cells in the upper respiratory tract . a) Poly (I:C) was formulated in DDA/MMG liposomes at a concentration of 12,500/2500/625 μg/mL DDA/MMG/Poly (I:C). This corresponds to a final dose of 250 μg/50 μg/12.5 μg DDA/MMG/Poly(I:C) for intranasal (i.n.) administration. The particle size (Z.ave) and polydispersity index (PDI) was measured using differential light scattering. b) A human TLR3 reporter (HEK293) cell line was used to assess delivery of Poly (I:C) in CAF09b. Cells were stimulated with Poly(I:C) alone (naked) or formulated in CAF09b at the indicated concentrations (100, 20, 4, and 0.8 μg/mL for naked Poly(I:C) and 1.67, 0.42, 0.1, 0.026, and 0.0065 μg/mL for Poly(I:C) in CAF09b). CAF04 (DDA/MMG without Poly(I:C)) was used as negative control. Secreted embryonic alkaline phosphatase produced upon TLR3 stimulation were determined using HEK-Blue™ Detection medium, measured as optical density (OD). c) The SARS-CoV-2 Spike HexaPro trimer was formulated in CAF09b adjuvant and confirmed to retain binding to ACE2 using ELISA. d) Mice were either left unvaccinated (naïve) or immunised with two doses of SARS-CoV-2 spike HexaPro trimer protein formulated in cationic liposomes (CAF®09b). The vaccine was administered as a conventional subcutaneous two dose regimen (s.c./s.c.), as subcutaneous priming followed by intranasal boosting (s.c./i.n.) or as two intranasal administrations (i.n./i.n.). Serum was sampled at 21 days after the 2nd immunisation. Created with BioRender.com . e) IgG antibody responses against the SARS-CoV-2 spike protein. f) Nasal wash IgA antibody responses against SARS-CoV-2 spike protein. Data are representative of n = 6 mice per group. Similar findings were obtained in another separate experiment ( e and f ). Spike-specific CD8 T cell responses were measured by flow cytometry. Mice were injected i.v. with anti-CD45.2 to distinguish between circulating and tissue resident cells. g) CD8 T cell responses in lungs and spleen. Cells were gated as CD45− (parenchymal cells not stained by the i.v., injected anti-CD45 antibody), CD8+ (to identify CD8 T cells), CD44+ (activation marker) SARS-CoV-2 spike-specific, using a tetramer (VNFNFNGL). Left panel shows representative stainings of lung cells. Data were pooled from two experiments with n = 6 mice and n = 9 mice, respectively. h) CD8 T cell responses in nasal-associated lymphoid tissue. The experiment was performed once and contained n = 9 mice. To obtain enough cells for analysis, each NALT data point was a pool from three mice. Mean ± SEM is displayed. Statistically significant differences are indicated by ∗ or ∗∗ (one-way ANOVA, comparing the mean of each column with the mean of the s.c./s.c. group, p < 0.05 or 0.01, respectively). There were no statistically significant differences among groups unless indicated.

Article Snippet: One million cells were treated with Fc-block (BD Biosciences) and stained in PBS +1% FBS with cocktails of antibodies against the following surface proteins: CD45.2 FITC (104, BD 553772), CD4 APC-Cy7 (RM4-5, 1:600, eBioscience 47-0042-82), CD4 BV786 (GK1.5, 1:200, BD 563331), CD8a PerCP-Cy5.5 (53-6.7, 1:600, eBioscience 45-0081-82), CD8a BV421 (53-6.7, 1:600, Biolegend 100738), CD44 APC (IM7, 1:600, BD 559250), CD44 APC-Cy7 (IM7, 1:200, eBioscience 47-0441-82), CD62L PerCP-Cy5.5 (MEL-14, 1:200, BD 560513), CD19 PE-Cy7 (1D3, 1:600, BD 552854), CD11c PE-Cy7 (HL3, 1:200, BD 558079), and I-A/I-E BV605 (M5/114.15.2, 1:200, BD 563413).

Techniques: Liposomes, Concentration Assay, Negative Control, Produced, Adjuvant, Binding Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Injection, Staining, Activation Assay, Marker

Journal: eBioMedicine

Article Title: Intranasal recombinant protein subunit vaccine targeting TLR3 induces respiratory tract IgA and CD8 T cell responses and protects against respiratory virus infection

doi: 10.1016/j.ebiom.2025.105615

Figure Lengend Snippet: Intranasal administration of CAF09b adjuvanted SARS-CoV-2 spike protein elicits higher respiratory tract immune responses than the licenced vaccine mRNA-1273 . Mice were immunised with two doses of SARS-CoV-2 spike HexaPro trimer (5 μg of protein) formulated in CAF09b adjuvant given intranasally (i.n./i.n.) or the licenced vaccine mRNA-1273 (Spikevax) vaccine (1 μg of mRNA given intramuscularly (i.m/i.m.)). a) Serum IgG antibody responses against spike protein. b) IgA antibody responses against spike protein measured in serum, lungs, nasal-associated lymphoid tissue (NALT), and nasal washes. Mice were injected i.v. with anti-CD45.2 to distinguish between circulating CD45+ (IV+) and tissue resident CD45− (IV−) cells c) CD8 T cell responses, as identified by gating on live+CD8+CD44+ cells binding a spike-specific tetramer (VNFNFNGL), were measured by flow cytometry. CD8 T cell responses were assessed systemically (IV+) in spleen (left panel), and locally (IV−) in lungs (middle panel) and NALT (right panel). Data represent two independent experiments with n = 3 (naïve) or 8–9 (vaccinated) mice per group. To obtain enough cells for analysis of CD8 T cell responses in NALT, each data point displays a pool of three mice. d) CD4 T cell responses, measured by gating for spike tetramer S62-76 (VTWFHAIHVSGTNGT) on live+CD8−CD19−CD4+CD62L−CD44+ cells, was assessed systemically (IV+) in spleen (left panel), and locally (IV−) in lungs (middle panel) and NALT (right panel). The experiment was performed once and data represent n = 3 (naïve) or n = 8–9 (vaccinated) mice per group. Mean ± SEM is displayed. Statistically significant differences are indicated by ∗, ∗∗, ∗∗∗ or ∗∗∗∗ (Student t-test, p < 0.05, 0.01, 0.001 or 0.0001, respectively). There were no statistically significant differences among groups unless indicated.

Article Snippet: One million cells were treated with Fc-block (BD Biosciences) and stained in PBS +1% FBS with cocktails of antibodies against the following surface proteins: CD45.2 FITC (104, BD 553772), CD4 APC-Cy7 (RM4-5, 1:600, eBioscience 47-0042-82), CD4 BV786 (GK1.5, 1:200, BD 563331), CD8a PerCP-Cy5.5 (53-6.7, 1:600, eBioscience 45-0081-82), CD8a BV421 (53-6.7, 1:600, Biolegend 100738), CD44 APC (IM7, 1:600, BD 559250), CD44 APC-Cy7 (IM7, 1:200, eBioscience 47-0441-82), CD62L PerCP-Cy5.5 (MEL-14, 1:200, BD 560513), CD19 PE-Cy7 (1D3, 1:600, BD 552854), CD11c PE-Cy7 (HL3, 1:200, BD 558079), and I-A/I-E BV605 (M5/114.15.2, 1:200, BD 563413).

Techniques: Adjuvant, Injection, Binding Assay, Flow Cytometry

Journal: Cell Death Discovery

Article Title: Endogenous osteoprotegerin (OPG) represses ERα and promotes stemness and chemoresistance in breast cancer cells

doi: 10.1038/s41420-024-02151-8

Figure Lengend Snippet: A Western blot images with GAPDH used as internal control. The numbers underneath each band represent fold changes relative to the control (Ctrl) after normalization to GAPDH. B Cells (2 × 10 5 ) were double‐stained for both CD24 and CD44, and the proportions of the subpopulation CD44 high /CD24 low were determined by flow cytometry and are shown in the boxes. C Cells (1 × 10 3 ) were seeded in ultra-low attachment 96‐well plate containing stem cell‐specific medium (100 μl). The number of spheroids (>100 μm) were counted. Representative photographs of spheroids (left panel), number of formed spheroids (right panel). Error bars represent mean ± SEM ( n = 3). * P ≤ 0.05; ** P ≤ 0.01. D Cells were plated in soft agar, and after 7 days, the number of colonies ( >100 μm) were counted. Representative photographs of colonies (left panel), number of formed colonies (right panel). Scale bars represent 100 μm. Error bars represent mean ± SEM ( n = 3). *** P ≤ 0.001. E Immunofluorescence, photographs were taken by fluorescent microscope. Scale bars represent 25 μm. F Different concentrations (2 × 10 3 , 2 × 10 4 , 2 × 10 5 , 2 × 10 6 , n = 3 for each cell concentration) of T-47D (OPG-ORF and Ctrl) cells were injected under the right and left nipple of female nude mice, respectively.

Article Snippet: Cells (2 × 10 5 ) were collected, washed, and incubated with CD44 APC-Cy7 (cat#559942) and CD24 PE (cat#555428) antibodies (BD biosciences) for surface staining (30 min on ice).

Techniques: Western Blot, Control, Staining, Flow Cytometry, Immunofluorescence, Microscopy, Concentration Assay, Injection

Journal: bioRxiv

Article Title: Rapid metabolic regulation of a novel arginine methylation of KCa3.1 attenuates T cell exhaustion

doi: 10.1101/2024.05.09.593421

Figure Lengend Snippet: (A) Quantification of intracellular Met in CD8 + T cells and B16 cells at 0, 6, and 12 hrs of co-culture (n=3). (B) Quantification of intracellular non-essential amino acids from LN CD8 + and CD8 + TIL, isolated at D12 post implantation of B16 tumors (n=3). (C) Quantification of intracellular Met from tumor cells and CD8 + TIL isolated from patient primary colorectal tumors (n=4). (D) T cell proliferation via cell-trace violet staining of OT-I CD8 + T cells activated in either 0.1 mM Met or 0.03 mM Met for the indicated times before restoration to 0.1 mM Met in 0.03 mM Met conditions, analyzed 72 hrs post activation (n=3). Representative of 3 samples per group. (E) Schematic design for OT-I T cell activation initially in 0.1 or 0.03 mM Met, followed by restoration of Met to 0.1 mM for 24 hrs before injection into B16-OVA tumor bearing mice. (F) Tumor growth of B16-OVA in Rag1 -/- treated with OT-I CD8 + T cells activated as described in (E) for 30 min-6hrs (n=5). (G) Schematic experimental design for generation of LCMV-GP33 specific memory T cells, which were then activated as in described in (E) before injection into B16-OVA tumor-bearing Rag1 -/- mice. (H, I) Tumor growth (h) and survival (i) of B16-OVA tumors treated as described in (G) (n=5). (J) Tumor growth and survival of B16-OVA tumor bearing WT mice treated with or without OT-I CD8 + T cells activated in 0.1 or 0.03 mM Met for 30 min before restoring Met to 0.1 mM in 0.03 mM Met for 24 hrs, as described in (E) (n=5). (K) Tumor growth of B16-OVA tumor bearing Rag1 -/- mice injected with OT-I T cells as in (E) (n=5). (L, M) Tumor weight (l) and frequency of CD62L + , Tcm (CD62L hi CD44 hi ), IFN-ψ + , and TOX expression (MFI) (m) in OT-I CD8 + TIL isolated from B16-OVA tumor bearing Rag1 -/- mice at D9 post-injection with OT-I T cells activated as in (E) (n=5). Data are mean±s.e.m. Unpaired two-tailed Student’s t-test (A, B, C, L and M), two-way ANOVA (F and H) and Mantel-Cox test (I and J-right). *P < 0.05, **P < 0.01, ****P < 0.0001

Article Snippet: BUV563 anti-mouse LAG3 (C9B7W, 741350), BUV615 anti-mouse CD69 (H1.2F3, 751593), BUV805 anti-mouse CXCR3 (173, 748700), BV421 anti-mouse EOMES (X4-83, 567166), BV480 anti-mouse CD45.1 (A20, 746666), Alexa-Fluor 488 anti-mouse TCF1 (S33-966, 567018), Alexa-Fluor 647 TOX (NAN448B, 568356), BUV 496 anti-mouse Ly-108 (13G3, 750046), APC/Cy7 anti-mouse CD44 (IM7, 560568) and BUV805 anti-mouse CD8α (53-6.7, 612898) were acquired from BD Biosciences.

Techniques: Co-Culture Assay, Isolation, Staining, Activation Assay, Injection, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Rapid metabolic regulation of a novel arginine methylation of KCa3.1 attenuates T cell exhaustion

doi: 10.1101/2024.05.09.593421

Figure Lengend Snippet: (A) Frequencies of T cells isolated from B16-OVA tumors at D12 post transfer of OT-I T cells activated as in (n=7). (B) Expression of TCF1 + on OT-I CD8 + T cells at D12 post transfer of OT-I T cells, activated as in (n=7). (C, D) Representative contour plot (c) and quantification (d) of TCF1 and Tim-3 expressing cells from TOX + CD8 + TIL from OT-I CD8 + T cells isolated at D12 post transfer of OT-I T cells activated as in into B16-OVA tumor-bearing Rag1 -/- mice (n=7). (E-G) Frequency of Tim-3 - Ly108 + TEXprog (e), Tim-3 + Ly108 - , CXCR3 - Tim-3 + TEXterm (f), CD62L + T memory and CD62L + CD44 + T central memory cells (g) in OT-I TIL as in (A) (n=7 (f), n=4 (g)). (H-J) Representative contour plot and quantification of CD127 + CD27 + memory cells (h, i) and KLRG1 + CD127 - terminal effector cells (j) in OT-I CD8 + TIL isolated at D12 post of cells activated as in into B16-OVA tumor-bearing Rag1 -/- mice (n=4). Data are mean±s.e.m with statistical analyses performed using paired two-tailed Student’s t-test (*P < 0.05, **P < 0.01, ***P < 0.001).

Article Snippet: BUV563 anti-mouse LAG3 (C9B7W, 741350), BUV615 anti-mouse CD69 (H1.2F3, 751593), BUV805 anti-mouse CXCR3 (173, 748700), BV421 anti-mouse EOMES (X4-83, 567166), BV480 anti-mouse CD45.1 (A20, 746666), Alexa-Fluor 488 anti-mouse TCF1 (S33-966, 567018), Alexa-Fluor 647 TOX (NAN448B, 568356), BUV 496 anti-mouse Ly-108 (13G3, 750046), APC/Cy7 anti-mouse CD44 (IM7, 560568) and BUV805 anti-mouse CD8α (53-6.7, 612898) were acquired from BD Biosciences.

Techniques: Isolation, Expressing, Two Tailed Test

Journal: bioRxiv

Article Title: Rapid metabolic regulation of a novel arginine methylation of KCa3.1 attenuates T cell exhaustion

doi: 10.1101/2024.05.09.593421

Figure Lengend Snippet: (A) Nuclear NFAT1 quantification in CD44 + CD8 + T cells isolated from B16 tumors and respective dLNs at D9 post implantation (each circle represents one cell, n=40 cells, n=5). (B) Quantification of nuclear NFAT1 in CD44 + CD8 + T cells isolated from B16 tumors, after 5 days of peri-tumoral supplementation per day of 50 μl HBSS or 61 μM Met (each circle represents one cell, n=40 cells, n=4). (C, D) Tumor growth (c) and survival (d) of B16 tumor-bearing WT mice, supplemented either with HBSS or 61 μM Met peri-tumorally for five days as in B (n=10). (E) B16 tumor growth (left) and survival (right) of WT mice, treated intraperitoneally with IgG isotype or anti-CD8 antibody at D-1 and D3 and treated peri-tumorally with HBSS or Met from D7-D12 as in B (n=5, representative of two experiments). (F) Schematic design to assess tumor growth with anti-PD1 treatment and with either HBSS or Met supplementation. (G, H) Tumour growth (g) and survival (h) of MC38 tumor-bearing WT mice, treated either with anti-PD1 or IgG-isotype and supplemented with HBSS or Met peri-tumorally for five days as in B (n=5, representative of two experiments). (I) Schematic of experimental design to assess the effect of HBSS or Met supplementation on CAR-T cell therapy in mouse solid tumors. (J, K) Tumor growth (j) and survival (k) of F420 tumor-bearing Rag1 -/- mice injected with B7-H3 CAR T-cells at D0 and peri-tumorally treated with HBSS or Met from D1-D6 as in B (n=5). Data are mean±s.e.m. Unpaired two-tailed Student’s t-test (A), 2-way ANOVA (C, E-left, G and J), and Mantel-Cox log rank test (D, E-right, H and K). Statistical analysis of (B) is described in methods. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.s

Article Snippet: BUV563 anti-mouse LAG3 (C9B7W, 741350), BUV615 anti-mouse CD69 (H1.2F3, 751593), BUV805 anti-mouse CXCR3 (173, 748700), BV421 anti-mouse EOMES (X4-83, 567166), BV480 anti-mouse CD45.1 (A20, 746666), Alexa-Fluor 488 anti-mouse TCF1 (S33-966, 567018), Alexa-Fluor 647 TOX (NAN448B, 568356), BUV 496 anti-mouse Ly-108 (13G3, 750046), APC/Cy7 anti-mouse CD44 (IM7, 560568) and BUV805 anti-mouse CD8α (53-6.7, 612898) were acquired from BD Biosciences.

Techniques: Isolation, Injection, Two Tailed Test

Journal: bioRxiv

Article Title: Rapid metabolic regulation of a novel arginine methylation of KCa3.1 attenuates T cell exhaustion

doi: 10.1101/2024.05.09.593421

Figure Lengend Snippet: (A) Schematic of experimental design of acute HBSS/Met treatment in sub-cutaneous tumors. (B) Nuclear NFAT1 quantification of CD44 + CD8 + T cells isolated from B16 subcutaneous tumors and dLN, 5 days post daily peri-tumoral supplementation of 50 μl 61μM Met (n=4 mice, each circle represents one cell, n=40 cells). (C-E) Tumor growth (left) and survival (right) of WT mice with sub-cutaneous MC38 colorectal tumors (c, n=10), Lewis-lung carcinoma (LLC) tumors (d, (representative of two experiments, n=5) and F420 osteosarcoma tumors (e, n=5), treated with either HBSS or 50 μl 61μM Met peri-tumorally daily for 5 days (n=10). (F, G) Expression of PD1, Tim-3, CD69 + (f), Tcm (CD62L hi Cd44 hi ) and Tem (CD62L lo CD44 hi ) (g) on CD8 + TIL isolated from B16 tumor bearing mice 2 days after 5 daily peri-tumoral injection with HBSS or Met (n=4). (H) Tumor growth of B16f10 and MC38 tumors in NSG mice treated peri-tumorally with 50 μl HBSS or 50 μl 61μM Met daily for 5 days (n=5). (I) Representative flow cytometry plot of blood showing CD8 + T cell depletion in anti-CD8 antibody treated mice compared to IgG-Isotype treated mice (n=4). (J) Schematic of experimental design of contralateral tumor experiment. (K) Growth of B16f10 tumors implanted on left shoulder (dashed lines) and right flank (solid lines) following peri-tumoral injection of flank tumor with 50 μl HBSS or 61μM Met daily for 5 days (n=5, representative of two experiments). (L) B16 tumor growth in WT mice fed with either 1 % Met chow or 1.5 % Met chow, 7 days post tumor implantation (n=5). Data are mean±s.e.m. Two-way ANOVA (C-F, J and K), Mantel-Cox log rank test (C, D and E), unpaired two-tailed Student’s t-test (B, G). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: BUV563 anti-mouse LAG3 (C9B7W, 741350), BUV615 anti-mouse CD69 (H1.2F3, 751593), BUV805 anti-mouse CXCR3 (173, 748700), BV421 anti-mouse EOMES (X4-83, 567166), BV480 anti-mouse CD45.1 (A20, 746666), Alexa-Fluor 488 anti-mouse TCF1 (S33-966, 567018), Alexa-Fluor 647 TOX (NAN448B, 568356), BUV 496 anti-mouse Ly-108 (13G3, 750046), APC/Cy7 anti-mouse CD44 (IM7, 560568) and BUV805 anti-mouse CD8α (53-6.7, 612898) were acquired from BD Biosciences.

Techniques: Isolation, Expressing, Injection, Flow Cytometry, Tumor Implantation, Two Tailed Test

Journal: Oncoimmunology

Article Title: The potential role of CMC1 as an immunometabolic checkpoint in T cell immunity

doi: 10.1080/2162402X.2024.2344905

Figure Lengend Snippet: Cmc1-/- CD8 + T cells retain more CD44 + CD62L + Tm cells and survive longer upon secondary activation in vitro.

Article Snippet: Antibodies: Alexa 700 anti-mouse CD25 (BioLegend; Cat# 102024); FITC anti-mouse IL-2 (BioLegend; Cat# 503806); FITC anti-mouse TNFα (BioLegend; Cat# 506304); BV605 anti-mouse IFNγ (BioLegend; Cat# 505840); Percp-cy5.5 anti-mouse GZMB (Biolegend; Cat# 372212); APC anti-mouse Perforin (BioLegend; Cat# 154403); APC anti-mouse EOMES (Invitrogen; Cat# 2473687); APC anti-mouse KLRG1 (BioLegend; Cat# 138412); BV421 anti-mouse CD127 (BioLegend; Cat# 135024); APC-Cy7 anti-mouse CD44 (BD Pharmingen; Cat# 560568); BV605 anti-mouse CD62L (BioLegend; Cat# 104437); PE-Cy7 anti-mouse CCR7 (BioLegend; Cat# 120123), BV421 anti-mouse PD-1 (Biolegend, Cat# 135221); PE-Cy7 anti-mouse Tim-3 (Biolegend, Cat# 134010); PE anti-mouse TOX (Invitrogen, Cat# 2407523); Alexa flour 647 anti-mouse TCF1 (BD Pharmingen, Cat# 566693).

Techniques: Activation Assay, In Vitro