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Immune cell profiling of tumor tissues from C57BL/6 mice injected with mEERL/sg-Scr or mEERL/sg2- Ulk1 (n=4/group). Isolated single cell suspensions were stained with an antibody cocktail and analyzed by flow cytometry. Percentages of total T cells of CD45 + (A), CD8 + T (B), and CD4 + T cells (C) of total CD3 + cells are shown. T cell profiling of tumor draining lymph nodes (TDLNs) from mice injected with either mEERL/sg-Scr or mEERL/sg- Ulk1 was isolated 21 days post-injection (n=3/group). Single-cell suspensions were stained with an antibody cocktail containing Zombie NIR, anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-CD44, and anti-CD62L antibodies. Representative bar graphs showing percentages of total T cells (D), CD4 + T (E), and CD8 + T cells (F), effector memory CD8 + T cells (G), and naïve CD8 + T cells (H). P -value was determined by One-way ANOVA. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. Cytolytic activity of isolated CD8 + T cells co-cultured with mEERL cells at a 50:1, 10:1, or 2:1 T cell: tumor cell ratios were measured by LDH assay (I). Effector functions of isolated CD8 + T cells were evaluated via IFNγ production measured by ELISA (J) and <t>ELISpot</t> (K and L). The number of spot-forming cells was counted using NIH ImageJ. All data shown are mean ± SEM and representative of at least 3 biological replicates. P -value was determined using Student’s t- Test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
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Immune cell profiling of tumor tissues from C57BL/6 mice injected with mEERL/sg-Scr or mEERL/sg2- Ulk1 (n=4/group). Isolated single cell suspensions were stained with an antibody cocktail and analyzed by flow cytometry. Percentages of total T cells of CD45 + (A), CD8 + T (B), and CD4 + T cells (C) of total CD3 + cells are shown. T cell profiling of tumor draining lymph nodes (TDLNs) from mice injected with either mEERL/sg-Scr or mEERL/sg- Ulk1 was isolated 21 days post-injection (n=3/group). Single-cell suspensions were stained with an antibody cocktail containing Zombie NIR, anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-CD44, and anti-CD62L antibodies. Representative bar graphs showing percentages of total T cells (D), CD4 + T (E), and CD8 + T cells (F), effector memory CD8 + T cells (G), and naïve CD8 + T cells (H). P -value was determined by One-way ANOVA. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. Cytolytic activity of isolated CD8 + T cells co-cultured with mEERL cells at a 50:1, 10:1, or 2:1 T cell: tumor cell ratios were measured by LDH assay (I). Effector functions of isolated CD8 + T cells were evaluated via IFNγ production measured by ELISA (J) and <t>ELISpot</t> (K and L). The number of spot-forming cells was counted using NIH ImageJ. All data shown are mean ± SEM and representative of at least 3 biological replicates. P -value was determined using Student’s t- Test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
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Immune cell profiling of tumor tissues from C57BL/6 mice injected with mEERL/sg-Scr or mEERL/sg2- Ulk1 (n=4/group). Isolated single cell suspensions were stained with an antibody cocktail and analyzed by flow cytometry. Percentages of total T cells of CD45 + (A), CD8 + T (B), and CD4 + T cells (C) of total CD3 + cells are shown. T cell profiling of tumor draining lymph nodes (TDLNs) from mice injected with either mEERL/sg-Scr or mEERL/sg- Ulk1 was isolated 21 days post-injection (n=3/group). Single-cell suspensions were stained with an antibody cocktail containing Zombie NIR, anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-CD44, and anti-CD62L antibodies. Representative bar graphs showing percentages of total T cells (D), CD4 + T (E), and CD8 + T cells (F), effector memory CD8 + T cells (G), and naïve CD8 + T cells (H). P -value was determined by One-way ANOVA. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. Cytolytic activity of isolated CD8 + T cells co-cultured with mEERL cells at a 50:1, 10:1, or 2:1 T cell: tumor cell ratios were measured by LDH assay (I). Effector functions of isolated CD8 + T cells were evaluated via IFNγ production measured by ELISA (J) and <t>ELISpot</t> (K and L). The number of spot-forming cells was counted using NIH ImageJ. All data shown are mean ± SEM and representative of at least 3 biological replicates. P -value was determined using Student’s t- Test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
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Immune cell profiling of tumor tissues from C57BL/6 mice injected with mEERL/sg-Scr or mEERL/sg2- Ulk1 (n=4/group). Isolated single cell suspensions were stained with an antibody cocktail and analyzed by flow cytometry. Percentages of total T cells of CD45 + (A), CD8 + T (B), and CD4 + T cells (C) of total CD3 + cells are shown. T cell profiling of tumor draining lymph nodes (TDLNs) from mice injected with either mEERL/sg-Scr or mEERL/sg- Ulk1 was isolated 21 days post-injection (n=3/group). Single-cell suspensions were stained with an antibody cocktail containing Zombie NIR, anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-CD44, and anti-CD62L antibodies. Representative bar graphs showing percentages of total T cells (D), CD4 + T (E), and CD8 + T cells (F), effector memory CD8 + T cells (G), and naïve CD8 + T cells (H). P -value was determined by One-way ANOVA. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. Cytolytic activity of isolated CD8 + T cells co-cultured with mEERL cells at a 50:1, 10:1, or 2:1 T cell: tumor cell ratios were measured by LDH assay (I). Effector functions of isolated CD8 + T cells were evaluated via IFNγ production measured by ELISA (J) and <t>ELISpot</t> (K and L). The number of spot-forming cells was counted using NIH ImageJ. All data shown are mean ± SEM and representative of at least 3 biological replicates. P -value was determined using Student’s t- Test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
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Immune cell profiling of tumor tissues from C57BL/6 mice injected with mEERL/sg-Scr or mEERL/sg2- Ulk1 (n=4/group). Isolated single cell suspensions were stained with an antibody cocktail and analyzed by flow cytometry. Percentages of total T cells of CD45 + (A), CD8 + T (B), and CD4 + T cells (C) of total CD3 + cells are shown. T cell profiling of tumor draining lymph nodes (TDLNs) from mice injected with either mEERL/sg-Scr or mEERL/sg- Ulk1 was isolated 21 days post-injection (n=3/group). Single-cell suspensions were stained with an antibody cocktail containing Zombie NIR, anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-CD44, and anti-CD62L antibodies. Representative bar graphs showing percentages of total T cells (D), CD4 + T (E), and CD8 + T cells (F), effector memory CD8 + T cells (G), and naïve CD8 + T cells (H). P -value was determined by One-way ANOVA. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. Cytolytic activity of isolated CD8 + T cells co-cultured with mEERL cells at a 50:1, 10:1, or 2:1 T cell: tumor cell ratios were measured by LDH assay (I). Effector functions of isolated CD8 + T cells were evaluated via IFNγ production measured by ELISA (J) and <t>ELISpot</t> (K and L). The number of spot-forming cells was counted using NIH ImageJ. All data shown are mean ± SEM and representative of at least 3 biological replicates. P -value was determined using Student’s t- Test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
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Immune cell profiling of tumor tissues from C57BL/6 mice injected with mEERL/sg-Scr or mEERL/sg2- Ulk1 (n=4/group). Isolated single cell suspensions were stained with an antibody cocktail and analyzed by flow cytometry. Percentages of total T cells of CD45 + (A), CD8 + T (B), and CD4 + T cells (C) of total CD3 + cells are shown. T cell profiling of tumor draining lymph nodes (TDLNs) from mice injected with either mEERL/sg-Scr or mEERL/sg- Ulk1 was isolated 21 days post-injection (n=3/group). Single-cell suspensions were stained with an antibody cocktail containing Zombie NIR, anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-CD44, and anti-CD62L antibodies. Representative bar graphs showing percentages of total T cells (D), CD4 + T (E), and CD8 + T cells (F), effector memory CD8 + T cells (G), and naïve CD8 + T cells (H). P -value was determined by One-way ANOVA. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. Cytolytic activity of isolated CD8 + T cells co-cultured with mEERL cells at a 50:1, 10:1, or 2:1 T cell: tumor cell ratios were measured by LDH assay (I). Effector functions of isolated CD8 + T cells were evaluated via IFNγ production measured by ELISA (J) and <t>ELISpot</t> (K and L). The number of spot-forming cells was counted using NIH ImageJ. All data shown are mean ± SEM and representative of at least 3 biological replicates. P -value was determined using Student’s t- Test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
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Immune cell profiling of tumor tissues from C57BL/6 mice injected with mEERL/sg-Scr or mEERL/sg2- Ulk1 (n=4/group). Isolated single cell suspensions were stained with an antibody cocktail and analyzed by flow cytometry. Percentages of total T cells of CD45 + (A), CD8 + T (B), and CD4 + T cells (C) of total CD3 + cells are shown. T cell profiling of tumor draining lymph nodes (TDLNs) from mice injected with either mEERL/sg-Scr or mEERL/sg- Ulk1 was isolated 21 days post-injection (n=3/group). Single-cell suspensions were stained with an antibody cocktail containing Zombie NIR, anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-CD44, and anti-CD62L antibodies. Representative bar graphs showing percentages of total T cells (D), CD4 + T (E), and CD8 + T cells (F), effector memory CD8 + T cells (G), and naïve CD8 + T cells (H). P -value was determined by One-way ANOVA. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. Cytolytic activity of isolated CD8 + T cells co-cultured with mEERL cells at a 50:1, 10:1, or 2:1 T cell: tumor cell ratios were measured by LDH assay (I). Effector functions of isolated CD8 + T cells were evaluated via IFNγ production measured by ELISA (J) and <t>ELISpot</t> (K and L). The number of spot-forming cells was counted using NIH ImageJ. All data shown are mean ± SEM and representative of at least 3 biological replicates. P -value was determined using Student’s t- Test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
Mouse Il 4 Elispotdevelopment Module 5 Plate, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Immune cell profiling of tumor tissues from C57BL/6 mice injected with mEERL/sg-Scr or mEERL/sg2- Ulk1 (n=4/group). Isolated single cell suspensions were stained with an antibody cocktail and analyzed by flow cytometry. Percentages of total T cells of CD45 + (A), CD8 + T (B), and CD4 + T cells (C) of total CD3 + cells are shown. T cell profiling of tumor draining lymph nodes (TDLNs) from mice injected with either mEERL/sg-Scr or mEERL/sg- Ulk1 was isolated 21 days post-injection (n=3/group). Single-cell suspensions were stained with an antibody cocktail containing Zombie NIR, anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-CD44, and anti-CD62L antibodies. Representative bar graphs showing percentages of total T cells (D), CD4 + T (E), and CD8 + T cells (F), effector memory CD8 + T cells (G), and naïve CD8 + T cells (H). P -value was determined by One-way ANOVA. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. Cytolytic activity of isolated CD8 + T cells co-cultured with mEERL cells at a 50:1, 10:1, or 2:1 T cell: tumor cell ratios were measured by LDH assay (I). Effector functions of isolated CD8 + T cells were evaluated via IFNγ production measured by ELISA (J) and <t>ELISpot</t> (K and L). The number of spot-forming cells was counted using NIH ImageJ. All data shown are mean ± SEM and representative of at least 3 biological replicates. P -value was determined using Student’s t- Test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
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Immune cell profiling of tumor tissues from C57BL/6 mice injected with mEERL/sg-Scr or mEERL/sg2- Ulk1 (n=4/group). Isolated single cell suspensions were stained with an antibody cocktail and analyzed by flow cytometry. Percentages of total T cells of CD45 + (A), CD8 + T (B), and CD4 + T cells (C) of total CD3 + cells are shown. T cell profiling of tumor draining lymph nodes (TDLNs) from mice injected with either mEERL/sg-Scr or mEERL/sg- Ulk1 was isolated 21 days post-injection (n=3/group). Single-cell suspensions were stained with an antibody cocktail containing Zombie NIR, anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-CD44, and anti-CD62L antibodies. Representative bar graphs showing percentages of total T cells (D), CD4 + T (E), and CD8 + T cells (F), effector memory CD8 + T cells (G), and naïve CD8 + T cells (H). P -value was determined by One-way ANOVA. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. Cytolytic activity of isolated CD8 + T cells co-cultured with mEERL cells at a 50:1, 10:1, or 2:1 T cell: tumor cell ratios were measured by LDH assay (I). Effector functions of isolated CD8 + T cells were evaluated via IFNγ production measured by ELISA (J) and <t>ELISpot</t> (K and L). The number of spot-forming cells was counted using NIH ImageJ. All data shown are mean ± SEM and representative of at least 3 biological replicates. P -value was determined using Student’s t- Test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.
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Immune cell profiling of tumor tissues from C57BL/6 mice injected with mEERL/sg-Scr or mEERL/sg2- Ulk1 (n=4/group). Isolated single cell suspensions were stained with an antibody cocktail and analyzed by flow cytometry. Percentages of total T cells of CD45 + (A), CD8 + T (B), and CD4 + T cells (C) of total CD3 + cells are shown. T cell profiling of tumor draining lymph nodes (TDLNs) from mice injected with either mEERL/sg-Scr or mEERL/sg- Ulk1 was isolated 21 days post-injection (n=3/group). Single-cell suspensions were stained with an antibody cocktail containing Zombie NIR, anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-CD44, and anti-CD62L antibodies. Representative bar graphs showing percentages of total T cells (D), CD4 + T (E), and CD8 + T cells (F), effector memory CD8 + T cells (G), and naïve CD8 + T cells (H). P -value was determined by One-way ANOVA. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. Cytolytic activity of isolated CD8 + T cells co-cultured with mEERL cells at a 50:1, 10:1, or 2:1 T cell: tumor cell ratios were measured by LDH assay (I). Effector functions of isolated CD8 + T cells were evaluated via IFNγ production measured by ELISA (J) and ELISpot (K and L). The number of spot-forming cells was counted using NIH ImageJ. All data shown are mean ± SEM and representative of at least 3 biological replicates. P -value was determined using Student’s t- Test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Journal: bioRxiv

Article Title: ULK1 drives NDP52-mediated selective autophagic degradation of MHC-I to promote immune evasion in HPV-positive head and neck cancer

doi: 10.64898/2026.03.14.711071

Figure Lengend Snippet: Immune cell profiling of tumor tissues from C57BL/6 mice injected with mEERL/sg-Scr or mEERL/sg2- Ulk1 (n=4/group). Isolated single cell suspensions were stained with an antibody cocktail and analyzed by flow cytometry. Percentages of total T cells of CD45 + (A), CD8 + T (B), and CD4 + T cells (C) of total CD3 + cells are shown. T cell profiling of tumor draining lymph nodes (TDLNs) from mice injected with either mEERL/sg-Scr or mEERL/sg- Ulk1 was isolated 21 days post-injection (n=3/group). Single-cell suspensions were stained with an antibody cocktail containing Zombie NIR, anti-CD45, anti-CD3, anti-CD4, anti-CD8, anti-CD44, and anti-CD62L antibodies. Representative bar graphs showing percentages of total T cells (D), CD4 + T (E), and CD8 + T cells (F), effector memory CD8 + T cells (G), and naïve CD8 + T cells (H). P -value was determined by One-way ANOVA. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001. Cytolytic activity of isolated CD8 + T cells co-cultured with mEERL cells at a 50:1, 10:1, or 2:1 T cell: tumor cell ratios were measured by LDH assay (I). Effector functions of isolated CD8 + T cells were evaluated via IFNγ production measured by ELISA (J) and ELISpot (K and L). The number of spot-forming cells was counted using NIH ImageJ. All data shown are mean ± SEM and representative of at least 3 biological replicates. P -value was determined using Student’s t- Test. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: IFN-γ production was determined using the Mouse IFN-γ ELISpot Development Module (R&D Systems, No. SEL485), visualized using the ELISpot Blue Color Module (Strep-AP and BCIP-BNBT) (R&D Systems, No. SEL002), and imaged using the ImmunoSpot Analyzer (CTL).

Techniques: Injection, Isolation, Single Cell, Staining, Flow Cytometry, Activity Assay, Cell Culture, Lactate Dehydrogenase Assay, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot