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b6 thy1 1 mice  (Jackson Laboratory)

 
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    Jackson Laboratory b6 thy1 1 mice
    A. Experimental scheme: naive TCR TAG <t>(Thy1.1)</t> were adoptively transferred into LM TAG -infected C57BL/6 (B6; Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers of infected mice (green) and tumor mice (blue) for flow cytometric analysis between 2.5-70+ days post-transfer. B. TCR TAG cell numbers in the spleens (left) and livers (right) of LM TAG -infected mice and tumor-bearing mice. Dots represent mean and error bars represent SEM. For B-D and F n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. C. Histograms (left) showing TCR TAG PD1 expression from infection shown in comparison to naive (N; gray). For C-D and F , each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. D. Histograms and summary plots of CD38 expression and CD101 expression in spleens and livers from tumor mice shown in comparison to N. E. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Right, summary plots of % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. *, P <0.05; **, P <0.01; ****, P <0.0001 (one-way ANOVA with post hoc Tukey test). F. Histograms and summary plots of CD122 expression and BCL2 expression from tumor mice shown in comparison to N. All flow plots are gated on live CD8 + Thy1.1 + cells, and flow data for each timepoint is concatenated from all biological replicates. Summary plots (right) show MFI (geometric m ean fluorescence intensity).
    B6 Thy1 1 Mice, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "TCF1 lo CD8 T cells proliferate and persist autonomously in tumors"

    Article Title: TCF1 lo CD8 T cells proliferate and persist autonomously in tumors

    Journal: bioRxiv

    doi: 10.64898/2026.01.17.700120

    A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into LM TAG -infected C57BL/6 (B6; Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers of infected mice (green) and tumor mice (blue) for flow cytometric analysis between 2.5-70+ days post-transfer. B. TCR TAG cell numbers in the spleens (left) and livers (right) of LM TAG -infected mice and tumor-bearing mice. Dots represent mean and error bars represent SEM. For B-D and F n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. C. Histograms (left) showing TCR TAG PD1 expression from infection shown in comparison to naive (N; gray). For C-D and F , each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. D. Histograms and summary plots of CD38 expression and CD101 expression in spleens and livers from tumor mice shown in comparison to N. E. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Right, summary plots of % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. *, P <0.05; **, P <0.01; ****, P <0.0001 (one-way ANOVA with post hoc Tukey test). F. Histograms and summary plots of CD122 expression and BCL2 expression from tumor mice shown in comparison to N. All flow plots are gated on live CD8 + Thy1.1 + cells, and flow data for each timepoint is concatenated from all biological replicates. Summary plots (right) show MFI (geometric m ean fluorescence intensity).
    Figure Legend Snippet: A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into LM TAG -infected C57BL/6 (B6; Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers of infected mice (green) and tumor mice (blue) for flow cytometric analysis between 2.5-70+ days post-transfer. B. TCR TAG cell numbers in the spleens (left) and livers (right) of LM TAG -infected mice and tumor-bearing mice. Dots represent mean and error bars represent SEM. For B-D and F n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. C. Histograms (left) showing TCR TAG PD1 expression from infection shown in comparison to naive (N; gray). For C-D and F , each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. D. Histograms and summary plots of CD38 expression and CD101 expression in spleens and livers from tumor mice shown in comparison to N. E. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Right, summary plots of % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. *, P <0.05; **, P <0.01; ****, P <0.0001 (one-way ANOVA with post hoc Tukey test). F. Histograms and summary plots of CD122 expression and BCL2 expression from tumor mice shown in comparison to N. All flow plots are gated on live CD8 + Thy1.1 + cells, and flow data for each timepoint is concatenated from all biological replicates. Summary plots (right) show MFI (geometric m ean fluorescence intensity).

    Techniques Used: Infection, Isolation, Expressing, Comparison, Ex Vivo, Fluorescence

    A. Experimental setup as shown in . Histograms and summary plots of CD44 expression from infection (green) and tumor (blue) in spleen and liver shown in comparison to naive (N; gray). Each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. B. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Summary plots show % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. ****, P <0.0001 (two-way ANOVA followed by post hoc Šídák’s multiple comparisons test). All flow plots are gated on live CD8 + Thy1.1 + cells and flow data for each timepoint is concatenated from all biological replicates. Data is representative of two independent experiments.
    Figure Legend Snippet: A. Experimental setup as shown in . Histograms and summary plots of CD44 expression from infection (green) and tumor (blue) in spleen and liver shown in comparison to naive (N; gray). Each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. B. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Summary plots show % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. ****, P <0.0001 (two-way ANOVA followed by post hoc Šídák’s multiple comparisons test). All flow plots are gated on live CD8 + Thy1.1 + cells and flow data for each timepoint is concatenated from all biological replicates. Data is representative of two independent experiments.

    Techniques Used: Expressing, Infection, Comparison, Ex Vivo

    A. Experimental setup as in . Heatmap summarizing data for flow cytometric assessment of activation and homing proteins, inhibitory receptors, cytokine receptors, proliferation and survival markers, and transcription factors. Blue-red color scale shows row normalized log 2 fold change (log2FC) of MFI in comparison to N. Each column represents individual biologic replicates for each condition and time point (the same single naive replicate is shown next to both infection and tumor groups). Data is representative of two independent experiments . B. Corresponding histograms showing expression of activation and homing markers (LY108, CD62L, CD44, CD69) andinhibitory receptors (PD1, CD38, CD101, CD101, and TIM3). C. Histograms showing expression of cytokine receptors (CD132, CD25, CD127, CD122), proliferation and survival markers (KI67, BCL2, BIM), and transcription factors (TOX, LEF1, and TCF1). All flow plots are gated on live CD8 + Thy1.1 + cells and flow data for each timepoint is concatenated from all biological replicates. Data is representative of two independent experiments.
    Figure Legend Snippet: A. Experimental setup as in . Heatmap summarizing data for flow cytometric assessment of activation and homing proteins, inhibitory receptors, cytokine receptors, proliferation and survival markers, and transcription factors. Blue-red color scale shows row normalized log 2 fold change (log2FC) of MFI in comparison to N. Each column represents individual biologic replicates for each condition and time point (the same single naive replicate is shown next to both infection and tumor groups). Data is representative of two independent experiments . B. Corresponding histograms showing expression of activation and homing markers (LY108, CD62L, CD44, CD69) andinhibitory receptors (PD1, CD38, CD101, CD101, and TIM3). C. Histograms showing expression of cytokine receptors (CD132, CD25, CD127, CD122), proliferation and survival markers (KI67, BCL2, BIM), and transcription factors (TOX, LEF1, and TCF1). All flow plots are gated on live CD8 + Thy1.1 + cells and flow data for each timepoint is concatenated from all biological replicates. Data is representative of two independent experiments.

    Techniques Used: Activation Assay, Comparison, Infection, Expressing

    A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers for flow cytometric analysis at 2.5, 5, 10, 14, 28, 35, and 70+ days post-transfer. B. Histograms and summary plots of KI67 expression of TCR TAG in spleens and livers from tumor-bearing mice shown in comparison to N, with % KI67 + gate set to exclude N. Flow plots for each time point were concatenated from all biological replicates; each symbol represents an individual mouse. *, P <0.007 determined by one-sample Student’s t -test with Bonferroni correction. C. Left, representative dot plots of KI67 expression and DNA content staining in TCR TAG in spleens and livers of AST;Cre-ER T2 mice, with inset numbers indicating the percentage of cells in G 0 , G 1 , and S-G 2 M phases of cell cycle. Middle, summary plots of the percentage of TCR TAG in G 0 , G 1 , and S-G 2 M, shown as mean ± SEM. Right, summary plots of the percentage of TCR TAG in S-G 2 M. Each symbol represents an individual mouse. *, P <0.007 determined by one-sample Student’s t -test with Bonferroni correction. D. Histograms and summary plots of TCF1 expression in spleens and livers, with TCF1 + gate shown. Flow plots for each time point were concatenated from all biological replicates; each symbol represents an individual mouse. *, P <0.007 determined by one-sample Student’s t -test with Bonferroni correction. E. Longitudinal analysis of the absolute cell numbers of TCR TAG cells (blue) in the spleens and livers of tumor-bearing mice and the % TCF1 + TCR TAG (yellow), shown as mean ± SEM. F. Summary plots of the % TCF1 + (filled bars) and TCF1 − (open bars) TCR TAG within the S-G 2 /M + subset, shown as mean ± SEM. Data is representative of two independent experiments with n = 3 mice per group
    Figure Legend Snippet: A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers for flow cytometric analysis at 2.5, 5, 10, 14, 28, 35, and 70+ days post-transfer. B. Histograms and summary plots of KI67 expression of TCR TAG in spleens and livers from tumor-bearing mice shown in comparison to N, with % KI67 + gate set to exclude N. Flow plots for each time point were concatenated from all biological replicates; each symbol represents an individual mouse. *, P <0.007 determined by one-sample Student’s t -test with Bonferroni correction. C. Left, representative dot plots of KI67 expression and DNA content staining in TCR TAG in spleens and livers of AST;Cre-ER T2 mice, with inset numbers indicating the percentage of cells in G 0 , G 1 , and S-G 2 M phases of cell cycle. Middle, summary plots of the percentage of TCR TAG in G 0 , G 1 , and S-G 2 M, shown as mean ± SEM. Right, summary plots of the percentage of TCR TAG in S-G 2 M. Each symbol represents an individual mouse. *, P <0.007 determined by one-sample Student’s t -test with Bonferroni correction. D. Histograms and summary plots of TCF1 expression in spleens and livers, with TCF1 + gate shown. Flow plots for each time point were concatenated from all biological replicates; each symbol represents an individual mouse. *, P <0.007 determined by one-sample Student’s t -test with Bonferroni correction. E. Longitudinal analysis of the absolute cell numbers of TCR TAG cells (blue) in the spleens and livers of tumor-bearing mice and the % TCF1 + TCR TAG (yellow), shown as mean ± SEM. F. Summary plots of the % TCF1 + (filled bars) and TCF1 − (open bars) TCR TAG within the S-G 2 /M + subset, shown as mean ± SEM. Data is representative of two independent experiments with n = 3 mice per group

    Techniques Used: Isolation, Expressing, Comparison, Staining

    A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into LM TAG -infected B6 (Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice and 5-ethynyl-2’-deoxyurdine (EdU) and 5-bromo-2’-deoxyuridine (BrdU) were administered on days 35 and 42, respectively. TCR TAG were re-isolated on day 43 from spleens of infected mice (green) and spleens and livers of tumor-bearing mice (blue) for flow cytometric analysis. B. Schematic representation of the expected distributions of EdU and BrdU incorporation for T cells proliferating in a progenitor-progeny hierarchy (left) or stochastically (right). C. Top, dot plots of TCR TAG EdU and BrdU incorporation in infected mice and tumor-bearing mice. Unlabeled cells shown in gray with inset numbers showing the percentage of nucleoside labeled cells in each gate. Bottom, summary plots of % EdU + (red), EdU + ,BrdU + (purple), and BrdU + (cyan) TCR TAG within each nucleoside labeled subset. Each symbol represents an individual mouse. **, P <0.01 determined by repeated measures, one-way ANOVA with post hoc Tukey test. D. Concatenated histogram of TCF1 expression in nucleoside-labeled TCR TAG and summary plot of the percentage of nucleoside-labeled TCR TAG expressing TCF1. Each symbol represents an individual mouse. *, P <0.025 determined by one-sample Student’s t -test with Bonferroni correction with n=4-5 mice per group and representative of two independent experiments. Flow plots show data concatenated from all biological replicates.
    Figure Legend Snippet: A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into LM TAG -infected B6 (Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice and 5-ethynyl-2’-deoxyurdine (EdU) and 5-bromo-2’-deoxyuridine (BrdU) were administered on days 35 and 42, respectively. TCR TAG were re-isolated on day 43 from spleens of infected mice (green) and spleens and livers of tumor-bearing mice (blue) for flow cytometric analysis. B. Schematic representation of the expected distributions of EdU and BrdU incorporation for T cells proliferating in a progenitor-progeny hierarchy (left) or stochastically (right). C. Top, dot plots of TCR TAG EdU and BrdU incorporation in infected mice and tumor-bearing mice. Unlabeled cells shown in gray with inset numbers showing the percentage of nucleoside labeled cells in each gate. Bottom, summary plots of % EdU + (red), EdU + ,BrdU + (purple), and BrdU + (cyan) TCR TAG within each nucleoside labeled subset. Each symbol represents an individual mouse. **, P <0.01 determined by repeated measures, one-way ANOVA with post hoc Tukey test. D. Concatenated histogram of TCF1 expression in nucleoside-labeled TCR TAG and summary plot of the percentage of nucleoside-labeled TCR TAG expressing TCF1. Each symbol represents an individual mouse. *, P <0.025 determined by one-sample Student’s t -test with Bonferroni correction with n=4-5 mice per group and representative of two independent experiments. Flow plots show data concatenated from all biological replicates.

    Techniques Used: Infection, Isolation, BrdU Incorporation Assay, Labeling, Expressing

    A. Histogram of TCF1 expression 48 hours post transfection of naive Cas9;TCR TAG with sgRNA transfected with non-targeting control sgRNA (NTC; light green) or TCF7 sgRNA (TCF1KO; dark teal), data representative of two independent experiments. B. Experimental scheme: NTC or TCF1KO TCR TAG (Thy1.1) were adoptively transferred into tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. EdU was administered in the final 3 days. TCR TAG were re-isolated on day 35 from the spleens and livers for flow cytometric analysis. C. NTC and TCF1KO TCR TAG cell numbers recovered from tumor-bearing mice. D. Representative dot plots of KI67 expression and EdU incorporation of NTC and TCF1KO TCR TAG cells, with gates set based on KI67 − /EdU − cells. E. Top, summary plots of the percentage (left) and absolute number (right) of EdU + NTC or TCF1KO TCR TAG . Bottom, summary plots of the percentage (left) and absolute number (right) of KI67 + or TCF1KO TCR TAG . For C and E , each symbol represents an individual mouse, with two-way ANOVA followed by post hoc Šídák’s multiple comparisons test with n=4-5 mice per group and representative of two independent experiments.
    Figure Legend Snippet: A. Histogram of TCF1 expression 48 hours post transfection of naive Cas9;TCR TAG with sgRNA transfected with non-targeting control sgRNA (NTC; light green) or TCF7 sgRNA (TCF1KO; dark teal), data representative of two independent experiments. B. Experimental scheme: NTC or TCF1KO TCR TAG (Thy1.1) were adoptively transferred into tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. EdU was administered in the final 3 days. TCR TAG were re-isolated on day 35 from the spleens and livers for flow cytometric analysis. C. NTC and TCF1KO TCR TAG cell numbers recovered from tumor-bearing mice. D. Representative dot plots of KI67 expression and EdU incorporation of NTC and TCF1KO TCR TAG cells, with gates set based on KI67 − /EdU − cells. E. Top, summary plots of the percentage (left) and absolute number (right) of EdU + NTC or TCF1KO TCR TAG . Bottom, summary plots of the percentage (left) and absolute number (right) of KI67 + or TCF1KO TCR TAG . For C and E , each symbol represents an individual mouse, with two-way ANOVA followed by post hoc Šídák’s multiple comparisons test with n=4-5 mice per group and representative of two independent experiments.

    Techniques Used: Expressing, Transfection, Control, Isolation

    A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. FTY720 treatment or vehicle was administered every other day beginning on day −1 for 6 days or day 28 for 3 weeks. EdU was administered for the final 3 days prior to harvest. TCR TAG were re-isolated on day 5 and day 49 from the livers for flow cytometric analysis. B. Representative histogram and summary plot of % CD3ε + cells in peripheral blood 24 hours after initiation of FTY720 (purple) or vehicle (black) treatment. ****, P <000.1 determined by unpaired, two-tailed Student’s t -test. C. TCR TAG cell numbers in the livers of vehicle- (open black bars) and FTY720 (purple bars)-treated mice. Each symbol represents an individual mouse, with ****, P <0.0001 determined by two-way ANOVA followed by post hoc Šídák’s multiple comparisons test. D. Representative histogram of EdU+ incorporation in TCR TAG T cells and summary plot of % EdU + from the livers of vehicle- and FTY720-treated AST;Cre-ER T2 mice. Each symbol represents an individual mouse, two-way ANOVA followed by post hoc Šídák’s multiple comparisons test. E. Left, representative dot plots of TCF1 and KI67 expression in EdU + TCR TAG from the livers of AST;CreER T2 mice at day 5 (top) and day 49 (bottom) following vehicle and FTY720 treatment. Right, summary plots of % TCF1 hi (top) and KI67 + (bottom) of EdU + TCR TAG . Each symbol represents an individual mouse, two-way ANOVA followed by post hoc Šídák’s multiple comparisons test. For B-E , n=4-5 mice per group and representative of two independent experiments.
    Figure Legend Snippet: A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. FTY720 treatment or vehicle was administered every other day beginning on day −1 for 6 days or day 28 for 3 weeks. EdU was administered for the final 3 days prior to harvest. TCR TAG were re-isolated on day 5 and day 49 from the livers for flow cytometric analysis. B. Representative histogram and summary plot of % CD3ε + cells in peripheral blood 24 hours after initiation of FTY720 (purple) or vehicle (black) treatment. ****, P <000.1 determined by unpaired, two-tailed Student’s t -test. C. TCR TAG cell numbers in the livers of vehicle- (open black bars) and FTY720 (purple bars)-treated mice. Each symbol represents an individual mouse, with ****, P <0.0001 determined by two-way ANOVA followed by post hoc Šídák’s multiple comparisons test. D. Representative histogram of EdU+ incorporation in TCR TAG T cells and summary plot of % EdU + from the livers of vehicle- and FTY720-treated AST;Cre-ER T2 mice. Each symbol represents an individual mouse, two-way ANOVA followed by post hoc Šídák’s multiple comparisons test. E. Left, representative dot plots of TCF1 and KI67 expression in EdU + TCR TAG from the livers of AST;CreER T2 mice at day 5 (top) and day 49 (bottom) following vehicle and FTY720 treatment. Right, summary plots of % TCF1 hi (top) and KI67 + (bottom) of EdU + TCR TAG . Each symbol represents an individual mouse, two-way ANOVA followed by post hoc Šídák’s multiple comparisons test. For B-E , n=4-5 mice per group and representative of two independent experiments.

    Techniques Used: Isolation, Two Tailed Test, Expressing



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    cby pl b6 thy1 a scrj balb c thy1 1 mice  (Jackson Laboratory)
    86
    Jackson Laboratory cby pl b6 thy1 a scrj balb c thy1 1 mice
    A. Experimental scheme: naive TCR TAG <t>(Thy1.1)</t> were adoptively transferred into LM TAG -infected C57BL/6 (B6; Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers of infected mice (green) and tumor mice (blue) for flow cytometric analysis between 2.5-70+ days post-transfer. B. TCR TAG cell numbers in the spleens (left) and livers (right) of LM TAG -infected mice and tumor-bearing mice. Dots represent mean and error bars represent SEM. For B-D and F n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. C. Histograms (left) showing TCR TAG PD1 expression from infection shown in comparison to naive (N; gray). For C-D and F , each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. D. Histograms and summary plots of CD38 expression and CD101 expression in spleens and livers from tumor mice shown in comparison to N. E. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Right, summary plots of % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. *, P <0.05; **, P <0.01; ****, P <0.0001 (one-way ANOVA with post hoc Tukey test). F. Histograms and summary plots of CD122 expression and BCL2 expression from tumor mice shown in comparison to N. All flow plots are gated on live CD8 + Thy1.1 + cells, and flow data for each timepoint is concatenated from all biological replicates. Summary plots (right) show MFI (geometric m ean fluorescence intensity).
    Cby Pl B6 Thy1 A Scrj Balb C Thy1 1 Mice, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pmel-1 t-cell receptor (tcr)-transgenic mice [b6.cg thy1 -tg(tcratcrb)8rest/j  (Jackson Laboratory)
    90
    Jackson Laboratory pmel-1 t-cell receptor (tcr)-transgenic mice [b6.cg thy1 -tg(tcratcrb)8rest/j
    A. Experimental scheme: naive TCR TAG <t>(Thy1.1)</t> were adoptively transferred into LM TAG -infected C57BL/6 (B6; Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers of infected mice (green) and tumor mice (blue) for flow cytometric analysis between 2.5-70+ days post-transfer. B. TCR TAG cell numbers in the spleens (left) and livers (right) of LM TAG -infected mice and tumor-bearing mice. Dots represent mean and error bars represent SEM. For B-D and F n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. C. Histograms (left) showing TCR TAG PD1 expression from infection shown in comparison to naive (N; gray). For C-D and F , each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. D. Histograms and summary plots of CD38 expression and CD101 expression in spleens and livers from tumor mice shown in comparison to N. E. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Right, summary plots of % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. *, P <0.05; **, P <0.01; ****, P <0.0001 (one-way ANOVA with post hoc Tukey test). F. Histograms and summary plots of CD122 expression and BCL2 expression from tumor mice shown in comparison to N. All flow plots are gated on live CD8 + Thy1.1 + cells, and flow data for each timepoint is concatenated from all biological replicates. Summary plots (right) show MFI (geometric m ean fluorescence intensity).
    Pmel 1 T Cell Receptor (Tcr) Transgenic Mice [B6.Cg Thy1 Tg(tcratcrb)8rest/J, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    thy1.1 pmel-1 transgenic mice (b6.cg- thy1 a /cy tg[tcratcrb]8rest/j)  (Jackson Laboratory)
    90
    Jackson Laboratory thy1.1 pmel-1 transgenic mice (b6.cg- thy1 a /cy tg[tcratcrb]8rest/j)
    A. Experimental scheme: naive TCR TAG <t>(Thy1.1)</t> were adoptively transferred into LM TAG -infected C57BL/6 (B6; Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers of infected mice (green) and tumor mice (blue) for flow cytometric analysis between 2.5-70+ days post-transfer. B. TCR TAG cell numbers in the spleens (left) and livers (right) of LM TAG -infected mice and tumor-bearing mice. Dots represent mean and error bars represent SEM. For B-D and F n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. C. Histograms (left) showing TCR TAG PD1 expression from infection shown in comparison to naive (N; gray). For C-D and F , each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. D. Histograms and summary plots of CD38 expression and CD101 expression in spleens and livers from tumor mice shown in comparison to N. E. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Right, summary plots of % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. *, P <0.05; **, P <0.01; ****, P <0.0001 (one-way ANOVA with post hoc Tukey test). F. Histograms and summary plots of CD122 expression and BCL2 expression from tumor mice shown in comparison to N. All flow plots are gated on live CD8 + Thy1.1 + cells, and flow data for each timepoint is concatenated from all biological replicates. Summary plots (right) show MFI (geometric m ean fluorescence intensity).
    Thy1.1 Pmel 1 Transgenic Mice (B6.Cg Thy1 A /Cy Tg[Tcratcrb]8rest/J), supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    transgenic pmel-1 mice b6.cg- thy1 a /cy tg(tcratcrb)8rest/j  (Jackson Laboratory)
    90
    Jackson Laboratory transgenic pmel-1 mice b6.cg- thy1 a /cy tg(tcratcrb)8rest/j
    A. Experimental scheme: naive TCR TAG <t>(Thy1.1)</t> were adoptively transferred into LM TAG -infected C57BL/6 (B6; Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers of infected mice (green) and tumor mice (blue) for flow cytometric analysis between 2.5-70+ days post-transfer. B. TCR TAG cell numbers in the spleens (left) and livers (right) of LM TAG -infected mice and tumor-bearing mice. Dots represent mean and error bars represent SEM. For B-D and F n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. C. Histograms (left) showing TCR TAG PD1 expression from infection shown in comparison to naive (N; gray). For C-D and F , each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. D. Histograms and summary plots of CD38 expression and CD101 expression in spleens and livers from tumor mice shown in comparison to N. E. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Right, summary plots of % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. *, P <0.05; **, P <0.01; ****, P <0.0001 (one-way ANOVA with post hoc Tukey test). F. Histograms and summary plots of CD122 expression and BCL2 expression from tumor mice shown in comparison to N. All flow plots are gated on live CD8 + Thy1.1 + cells, and flow data for each timepoint is concatenated from all biological replicates. Summary plots (right) show MFI (geometric m ean fluorescence intensity).
    Transgenic Pmel 1 Mice B6.Cg Thy1 A /Cy Tg(tcratcrb)8rest/J, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    t cell receptor tcr transgenic thy1 1 pmel 1 pmel mice b6 cg thy1 a cy tg tcratcrb 8rest j  (Jackson Laboratory)
    86
    Jackson Laboratory t cell receptor tcr transgenic thy1 1 pmel 1 pmel mice b6 cg thy1 a cy tg tcratcrb 8rest j
    A. Experimental scheme: naive TCR TAG <t>(Thy1.1)</t> were adoptively transferred into LM TAG -infected C57BL/6 (B6; Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers of infected mice (green) and tumor mice (blue) for flow cytometric analysis between 2.5-70+ days post-transfer. B. TCR TAG cell numbers in the spleens (left) and livers (right) of LM TAG -infected mice and tumor-bearing mice. Dots represent mean and error bars represent SEM. For B-D and F n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. C. Histograms (left) showing TCR TAG PD1 expression from infection shown in comparison to naive (N; gray). For C-D and F , each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. D. Histograms and summary plots of CD38 expression and CD101 expression in spleens and livers from tumor mice shown in comparison to N. E. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Right, summary plots of % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. *, P <0.05; **, P <0.01; ****, P <0.0001 (one-way ANOVA with post hoc Tukey test). F. Histograms and summary plots of CD122 expression and BCL2 expression from tumor mice shown in comparison to N. All flow plots are gated on live CD8 + Thy1.1 + cells, and flow data for each timepoint is concatenated from all biological replicates. Summary plots (right) show MFI (geometric m ean fluorescence intensity).
    T Cell Receptor Tcr Transgenic Thy1 1 Pmel 1 Pmel Mice B6 Cg Thy1 A Cy Tg Tcratcrb 8rest J, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into LM TAG -infected C57BL/6 (B6; Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers of infected mice (green) and tumor mice (blue) for flow cytometric analysis between 2.5-70+ days post-transfer. B. TCR TAG cell numbers in the spleens (left) and livers (right) of LM TAG -infected mice and tumor-bearing mice. Dots represent mean and error bars represent SEM. For B-D and F n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. C. Histograms (left) showing TCR TAG PD1 expression from infection shown in comparison to naive (N; gray). For C-D and F , each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. D. Histograms and summary plots of CD38 expression and CD101 expression in spleens and livers from tumor mice shown in comparison to N. E. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Right, summary plots of % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. *, P <0.05; **, P <0.01; ****, P <0.0001 (one-way ANOVA with post hoc Tukey test). F. Histograms and summary plots of CD122 expression and BCL2 expression from tumor mice shown in comparison to N. All flow plots are gated on live CD8 + Thy1.1 + cells, and flow data for each timepoint is concatenated from all biological replicates. Summary plots (right) show MFI (geometric m ean fluorescence intensity).

    Journal: bioRxiv

    Article Title: TCF1 lo CD8 T cells proliferate and persist autonomously in tumors

    doi: 10.64898/2026.01.17.700120

    Figure Lengend Snippet: A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into LM TAG -infected C57BL/6 (B6; Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers of infected mice (green) and tumor mice (blue) for flow cytometric analysis between 2.5-70+ days post-transfer. B. TCR TAG cell numbers in the spleens (left) and livers (right) of LM TAG -infected mice and tumor-bearing mice. Dots represent mean and error bars represent SEM. For B-D and F n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. C. Histograms (left) showing TCR TAG PD1 expression from infection shown in comparison to naive (N; gray). For C-D and F , each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. D. Histograms and summary plots of CD38 expression and CD101 expression in spleens and livers from tumor mice shown in comparison to N. E. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Right, summary plots of % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. *, P <0.05; **, P <0.01; ****, P <0.0001 (one-way ANOVA with post hoc Tukey test). F. Histograms and summary plots of CD122 expression and BCL2 expression from tumor mice shown in comparison to N. All flow plots are gated on live CD8 + Thy1.1 + cells, and flow data for each timepoint is concatenated from all biological replicates. Summary plots (right) show MFI (geometric m ean fluorescence intensity).

    Article Snippet: TCR TAG transgenic mice (B6.Cg-Tg(TcraY1,TcrbY1)416Tev/J, RRID:IMSR_JAX:005236) , Cre-ER T2 mice (B6.129-Gt(ROSA)26Sortm1(cre/ERT2)Tyj/J, RRID:IMSR_JAX:008463), Rosa26-Cas9 mice (Gt(ROSA)26Sortm1.1(CAG-cas9*,-EGFP)Fezh/J, RRID:IMSR_JAX:026179), B6 Thy1.1 mice (B6.PL-Thy1a/CyJ, RRID:IMSR_JAX:000406), and B6 mice (C57BL/6J, RRID:IMSR_JAX:000664) were purchased from the Jackson Laboratory.

    Techniques: Infection, Isolation, Expressing, Comparison, Ex Vivo, Fluorescence

    A. Experimental setup as shown in . Histograms and summary plots of CD44 expression from infection (green) and tumor (blue) in spleen and liver shown in comparison to naive (N; gray). Each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. B. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Summary plots show % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. ****, P <0.0001 (two-way ANOVA followed by post hoc Šídák’s multiple comparisons test). All flow plots are gated on live CD8 + Thy1.1 + cells and flow data for each timepoint is concatenated from all biological replicates. Data is representative of two independent experiments.

    Journal: bioRxiv

    Article Title: TCF1 lo CD8 T cells proliferate and persist autonomously in tumors

    doi: 10.64898/2026.01.17.700120

    Figure Lengend Snippet: A. Experimental setup as shown in . Histograms and summary plots of CD44 expression from infection (green) and tumor (blue) in spleen and liver shown in comparison to naive (N; gray). Each symbol represents an individual mouse, and one-sample Student’s t -test with Bonferroni correction was used to determine significance: *, P <0.008 (infection); *, P <0.007 (tumor); ns, not statistically significant. n = 2-3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. B. TCR TAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Summary plots show % IFNγ + TCR TAG after peptide stimulation; each symbol represents an individual mouse; n = 3-4 mice per group. ****, P <0.0001 (two-way ANOVA followed by post hoc Šídák’s multiple comparisons test). All flow plots are gated on live CD8 + Thy1.1 + cells and flow data for each timepoint is concatenated from all biological replicates. Data is representative of two independent experiments.

    Article Snippet: TCR TAG transgenic mice (B6.Cg-Tg(TcraY1,TcrbY1)416Tev/J, RRID:IMSR_JAX:005236) , Cre-ER T2 mice (B6.129-Gt(ROSA)26Sortm1(cre/ERT2)Tyj/J, RRID:IMSR_JAX:008463), Rosa26-Cas9 mice (Gt(ROSA)26Sortm1.1(CAG-cas9*,-EGFP)Fezh/J, RRID:IMSR_JAX:026179), B6 Thy1.1 mice (B6.PL-Thy1a/CyJ, RRID:IMSR_JAX:000406), and B6 mice (C57BL/6J, RRID:IMSR_JAX:000664) were purchased from the Jackson Laboratory.

    Techniques: Expressing, Infection, Comparison, Ex Vivo

    A. Experimental setup as in . Heatmap summarizing data for flow cytometric assessment of activation and homing proteins, inhibitory receptors, cytokine receptors, proliferation and survival markers, and transcription factors. Blue-red color scale shows row normalized log 2 fold change (log2FC) of MFI in comparison to N. Each column represents individual biologic replicates for each condition and time point (the same single naive replicate is shown next to both infection and tumor groups). Data is representative of two independent experiments . B. Corresponding histograms showing expression of activation and homing markers (LY108, CD62L, CD44, CD69) andinhibitory receptors (PD1, CD38, CD101, CD101, and TIM3). C. Histograms showing expression of cytokine receptors (CD132, CD25, CD127, CD122), proliferation and survival markers (KI67, BCL2, BIM), and transcription factors (TOX, LEF1, and TCF1). All flow plots are gated on live CD8 + Thy1.1 + cells and flow data for each timepoint is concatenated from all biological replicates. Data is representative of two independent experiments.

    Journal: bioRxiv

    Article Title: TCF1 lo CD8 T cells proliferate and persist autonomously in tumors

    doi: 10.64898/2026.01.17.700120

    Figure Lengend Snippet: A. Experimental setup as in . Heatmap summarizing data for flow cytometric assessment of activation and homing proteins, inhibitory receptors, cytokine receptors, proliferation and survival markers, and transcription factors. Blue-red color scale shows row normalized log 2 fold change (log2FC) of MFI in comparison to N. Each column represents individual biologic replicates for each condition and time point (the same single naive replicate is shown next to both infection and tumor groups). Data is representative of two independent experiments . B. Corresponding histograms showing expression of activation and homing markers (LY108, CD62L, CD44, CD69) andinhibitory receptors (PD1, CD38, CD101, CD101, and TIM3). C. Histograms showing expression of cytokine receptors (CD132, CD25, CD127, CD122), proliferation and survival markers (KI67, BCL2, BIM), and transcription factors (TOX, LEF1, and TCF1). All flow plots are gated on live CD8 + Thy1.1 + cells and flow data for each timepoint is concatenated from all biological replicates. Data is representative of two independent experiments.

    Article Snippet: TCR TAG transgenic mice (B6.Cg-Tg(TcraY1,TcrbY1)416Tev/J, RRID:IMSR_JAX:005236) , Cre-ER T2 mice (B6.129-Gt(ROSA)26Sortm1(cre/ERT2)Tyj/J, RRID:IMSR_JAX:008463), Rosa26-Cas9 mice (Gt(ROSA)26Sortm1.1(CAG-cas9*,-EGFP)Fezh/J, RRID:IMSR_JAX:026179), B6 Thy1.1 mice (B6.PL-Thy1a/CyJ, RRID:IMSR_JAX:000406), and B6 mice (C57BL/6J, RRID:IMSR_JAX:000664) were purchased from the Jackson Laboratory.

    Techniques: Activation Assay, Comparison, Infection, Expressing

    A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers for flow cytometric analysis at 2.5, 5, 10, 14, 28, 35, and 70+ days post-transfer. B. Histograms and summary plots of KI67 expression of TCR TAG in spleens and livers from tumor-bearing mice shown in comparison to N, with % KI67 + gate set to exclude N. Flow plots for each time point were concatenated from all biological replicates; each symbol represents an individual mouse. *, P <0.007 determined by one-sample Student’s t -test with Bonferroni correction. C. Left, representative dot plots of KI67 expression and DNA content staining in TCR TAG in spleens and livers of AST;Cre-ER T2 mice, with inset numbers indicating the percentage of cells in G 0 , G 1 , and S-G 2 M phases of cell cycle. Middle, summary plots of the percentage of TCR TAG in G 0 , G 1 , and S-G 2 M, shown as mean ± SEM. Right, summary plots of the percentage of TCR TAG in S-G 2 M. Each symbol represents an individual mouse. *, P <0.007 determined by one-sample Student’s t -test with Bonferroni correction. D. Histograms and summary plots of TCF1 expression in spleens and livers, with TCF1 + gate shown. Flow plots for each time point were concatenated from all biological replicates; each symbol represents an individual mouse. *, P <0.007 determined by one-sample Student’s t -test with Bonferroni correction. E. Longitudinal analysis of the absolute cell numbers of TCR TAG cells (blue) in the spleens and livers of tumor-bearing mice and the % TCF1 + TCR TAG (yellow), shown as mean ± SEM. F. Summary plots of the % TCF1 + (filled bars) and TCF1 − (open bars) TCR TAG within the S-G 2 /M + subset, shown as mean ± SEM. Data is representative of two independent experiments with n = 3 mice per group

    Journal: bioRxiv

    Article Title: TCF1 lo CD8 T cells proliferate and persist autonomously in tumors

    doi: 10.64898/2026.01.17.700120

    Figure Lengend Snippet: A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. TCR TAG were re-isolated from spleens and livers for flow cytometric analysis at 2.5, 5, 10, 14, 28, 35, and 70+ days post-transfer. B. Histograms and summary plots of KI67 expression of TCR TAG in spleens and livers from tumor-bearing mice shown in comparison to N, with % KI67 + gate set to exclude N. Flow plots for each time point were concatenated from all biological replicates; each symbol represents an individual mouse. *, P <0.007 determined by one-sample Student’s t -test with Bonferroni correction. C. Left, representative dot plots of KI67 expression and DNA content staining in TCR TAG in spleens and livers of AST;Cre-ER T2 mice, with inset numbers indicating the percentage of cells in G 0 , G 1 , and S-G 2 M phases of cell cycle. Middle, summary plots of the percentage of TCR TAG in G 0 , G 1 , and S-G 2 M, shown as mean ± SEM. Right, summary plots of the percentage of TCR TAG in S-G 2 M. Each symbol represents an individual mouse. *, P <0.007 determined by one-sample Student’s t -test with Bonferroni correction. D. Histograms and summary plots of TCF1 expression in spleens and livers, with TCF1 + gate shown. Flow plots for each time point were concatenated from all biological replicates; each symbol represents an individual mouse. *, P <0.007 determined by one-sample Student’s t -test with Bonferroni correction. E. Longitudinal analysis of the absolute cell numbers of TCR TAG cells (blue) in the spleens and livers of tumor-bearing mice and the % TCF1 + TCR TAG (yellow), shown as mean ± SEM. F. Summary plots of the % TCF1 + (filled bars) and TCF1 − (open bars) TCR TAG within the S-G 2 /M + subset, shown as mean ± SEM. Data is representative of two independent experiments with n = 3 mice per group

    Article Snippet: TCR TAG transgenic mice (B6.Cg-Tg(TcraY1,TcrbY1)416Tev/J, RRID:IMSR_JAX:005236) , Cre-ER T2 mice (B6.129-Gt(ROSA)26Sortm1(cre/ERT2)Tyj/J, RRID:IMSR_JAX:008463), Rosa26-Cas9 mice (Gt(ROSA)26Sortm1.1(CAG-cas9*,-EGFP)Fezh/J, RRID:IMSR_JAX:026179), B6 Thy1.1 mice (B6.PL-Thy1a/CyJ, RRID:IMSR_JAX:000406), and B6 mice (C57BL/6J, RRID:IMSR_JAX:000664) were purchased from the Jackson Laboratory.

    Techniques: Isolation, Expressing, Comparison, Staining

    A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into LM TAG -infected B6 (Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice and 5-ethynyl-2’-deoxyurdine (EdU) and 5-bromo-2’-deoxyuridine (BrdU) were administered on days 35 and 42, respectively. TCR TAG were re-isolated on day 43 from spleens of infected mice (green) and spleens and livers of tumor-bearing mice (blue) for flow cytometric analysis. B. Schematic representation of the expected distributions of EdU and BrdU incorporation for T cells proliferating in a progenitor-progeny hierarchy (left) or stochastically (right). C. Top, dot plots of TCR TAG EdU and BrdU incorporation in infected mice and tumor-bearing mice. Unlabeled cells shown in gray with inset numbers showing the percentage of nucleoside labeled cells in each gate. Bottom, summary plots of % EdU + (red), EdU + ,BrdU + (purple), and BrdU + (cyan) TCR TAG within each nucleoside labeled subset. Each symbol represents an individual mouse. **, P <0.01 determined by repeated measures, one-way ANOVA with post hoc Tukey test. D. Concatenated histogram of TCF1 expression in nucleoside-labeled TCR TAG and summary plot of the percentage of nucleoside-labeled TCR TAG expressing TCF1. Each symbol represents an individual mouse. *, P <0.025 determined by one-sample Student’s t -test with Bonferroni correction with n=4-5 mice per group and representative of two independent experiments. Flow plots show data concatenated from all biological replicates.

    Journal: bioRxiv

    Article Title: TCF1 lo CD8 T cells proliferate and persist autonomously in tumors

    doi: 10.64898/2026.01.17.700120

    Figure Lengend Snippet: A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into LM TAG -infected B6 (Thy1.2) or tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice and 5-ethynyl-2’-deoxyurdine (EdU) and 5-bromo-2’-deoxyuridine (BrdU) were administered on days 35 and 42, respectively. TCR TAG were re-isolated on day 43 from spleens of infected mice (green) and spleens and livers of tumor-bearing mice (blue) for flow cytometric analysis. B. Schematic representation of the expected distributions of EdU and BrdU incorporation for T cells proliferating in a progenitor-progeny hierarchy (left) or stochastically (right). C. Top, dot plots of TCR TAG EdU and BrdU incorporation in infected mice and tumor-bearing mice. Unlabeled cells shown in gray with inset numbers showing the percentage of nucleoside labeled cells in each gate. Bottom, summary plots of % EdU + (red), EdU + ,BrdU + (purple), and BrdU + (cyan) TCR TAG within each nucleoside labeled subset. Each symbol represents an individual mouse. **, P <0.01 determined by repeated measures, one-way ANOVA with post hoc Tukey test. D. Concatenated histogram of TCF1 expression in nucleoside-labeled TCR TAG and summary plot of the percentage of nucleoside-labeled TCR TAG expressing TCF1. Each symbol represents an individual mouse. *, P <0.025 determined by one-sample Student’s t -test with Bonferroni correction with n=4-5 mice per group and representative of two independent experiments. Flow plots show data concatenated from all biological replicates.

    Article Snippet: TCR TAG transgenic mice (B6.Cg-Tg(TcraY1,TcrbY1)416Tev/J, RRID:IMSR_JAX:005236) , Cre-ER T2 mice (B6.129-Gt(ROSA)26Sortm1(cre/ERT2)Tyj/J, RRID:IMSR_JAX:008463), Rosa26-Cas9 mice (Gt(ROSA)26Sortm1.1(CAG-cas9*,-EGFP)Fezh/J, RRID:IMSR_JAX:026179), B6 Thy1.1 mice (B6.PL-Thy1a/CyJ, RRID:IMSR_JAX:000406), and B6 mice (C57BL/6J, RRID:IMSR_JAX:000664) were purchased from the Jackson Laboratory.

    Techniques: Infection, Isolation, BrdU Incorporation Assay, Labeling, Expressing

    A. Histogram of TCF1 expression 48 hours post transfection of naive Cas9;TCR TAG with sgRNA transfected with non-targeting control sgRNA (NTC; light green) or TCF7 sgRNA (TCF1KO; dark teal), data representative of two independent experiments. B. Experimental scheme: NTC or TCF1KO TCR TAG (Thy1.1) were adoptively transferred into tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. EdU was administered in the final 3 days. TCR TAG were re-isolated on day 35 from the spleens and livers for flow cytometric analysis. C. NTC and TCF1KO TCR TAG cell numbers recovered from tumor-bearing mice. D. Representative dot plots of KI67 expression and EdU incorporation of NTC and TCF1KO TCR TAG cells, with gates set based on KI67 − /EdU − cells. E. Top, summary plots of the percentage (left) and absolute number (right) of EdU + NTC or TCF1KO TCR TAG . Bottom, summary plots of the percentage (left) and absolute number (right) of KI67 + or TCF1KO TCR TAG . For C and E , each symbol represents an individual mouse, with two-way ANOVA followed by post hoc Šídák’s multiple comparisons test with n=4-5 mice per group and representative of two independent experiments.

    Journal: bioRxiv

    Article Title: TCF1 lo CD8 T cells proliferate and persist autonomously in tumors

    doi: 10.64898/2026.01.17.700120

    Figure Lengend Snippet: A. Histogram of TCF1 expression 48 hours post transfection of naive Cas9;TCR TAG with sgRNA transfected with non-targeting control sgRNA (NTC; light green) or TCF7 sgRNA (TCF1KO; dark teal), data representative of two independent experiments. B. Experimental scheme: NTC or TCF1KO TCR TAG (Thy1.1) were adoptively transferred into tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. EdU was administered in the final 3 days. TCR TAG were re-isolated on day 35 from the spleens and livers for flow cytometric analysis. C. NTC and TCF1KO TCR TAG cell numbers recovered from tumor-bearing mice. D. Representative dot plots of KI67 expression and EdU incorporation of NTC and TCF1KO TCR TAG cells, with gates set based on KI67 − /EdU − cells. E. Top, summary plots of the percentage (left) and absolute number (right) of EdU + NTC or TCF1KO TCR TAG . Bottom, summary plots of the percentage (left) and absolute number (right) of KI67 + or TCF1KO TCR TAG . For C and E , each symbol represents an individual mouse, with two-way ANOVA followed by post hoc Šídák’s multiple comparisons test with n=4-5 mice per group and representative of two independent experiments.

    Article Snippet: TCR TAG transgenic mice (B6.Cg-Tg(TcraY1,TcrbY1)416Tev/J, RRID:IMSR_JAX:005236) , Cre-ER T2 mice (B6.129-Gt(ROSA)26Sortm1(cre/ERT2)Tyj/J, RRID:IMSR_JAX:008463), Rosa26-Cas9 mice (Gt(ROSA)26Sortm1.1(CAG-cas9*,-EGFP)Fezh/J, RRID:IMSR_JAX:026179), B6 Thy1.1 mice (B6.PL-Thy1a/CyJ, RRID:IMSR_JAX:000406), and B6 mice (C57BL/6J, RRID:IMSR_JAX:000664) were purchased from the Jackson Laboratory.

    Techniques: Expressing, Transfection, Control, Isolation

    A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. FTY720 treatment or vehicle was administered every other day beginning on day −1 for 6 days or day 28 for 3 weeks. EdU was administered for the final 3 days prior to harvest. TCR TAG were re-isolated on day 5 and day 49 from the livers for flow cytometric analysis. B. Representative histogram and summary plot of % CD3ε + cells in peripheral blood 24 hours after initiation of FTY720 (purple) or vehicle (black) treatment. ****, P <000.1 determined by unpaired, two-tailed Student’s t -test. C. TCR TAG cell numbers in the livers of vehicle- (open black bars) and FTY720 (purple bars)-treated mice. Each symbol represents an individual mouse, with ****, P <0.0001 determined by two-way ANOVA followed by post hoc Šídák’s multiple comparisons test. D. Representative histogram of EdU+ incorporation in TCR TAG T cells and summary plot of % EdU + from the livers of vehicle- and FTY720-treated AST;Cre-ER T2 mice. Each symbol represents an individual mouse, two-way ANOVA followed by post hoc Šídák’s multiple comparisons test. E. Left, representative dot plots of TCF1 and KI67 expression in EdU + TCR TAG from the livers of AST;CreER T2 mice at day 5 (top) and day 49 (bottom) following vehicle and FTY720 treatment. Right, summary plots of % TCF1 hi (top) and KI67 + (bottom) of EdU + TCR TAG . Each symbol represents an individual mouse, two-way ANOVA followed by post hoc Šídák’s multiple comparisons test. For B-E , n=4-5 mice per group and representative of two independent experiments.

    Journal: bioRxiv

    Article Title: TCF1 lo CD8 T cells proliferate and persist autonomously in tumors

    doi: 10.64898/2026.01.17.700120

    Figure Lengend Snippet: A. Experimental scheme: naive TCR TAG (Thy1.1) were adoptively transferred into tamoxifen-treated AST;Cre-ER T2 (Thy1.2) mice. FTY720 treatment or vehicle was administered every other day beginning on day −1 for 6 days or day 28 for 3 weeks. EdU was administered for the final 3 days prior to harvest. TCR TAG were re-isolated on day 5 and day 49 from the livers for flow cytometric analysis. B. Representative histogram and summary plot of % CD3ε + cells in peripheral blood 24 hours after initiation of FTY720 (purple) or vehicle (black) treatment. ****, P <000.1 determined by unpaired, two-tailed Student’s t -test. C. TCR TAG cell numbers in the livers of vehicle- (open black bars) and FTY720 (purple bars)-treated mice. Each symbol represents an individual mouse, with ****, P <0.0001 determined by two-way ANOVA followed by post hoc Šídák’s multiple comparisons test. D. Representative histogram of EdU+ incorporation in TCR TAG T cells and summary plot of % EdU + from the livers of vehicle- and FTY720-treated AST;Cre-ER T2 mice. Each symbol represents an individual mouse, two-way ANOVA followed by post hoc Šídák’s multiple comparisons test. E. Left, representative dot plots of TCF1 and KI67 expression in EdU + TCR TAG from the livers of AST;CreER T2 mice at day 5 (top) and day 49 (bottom) following vehicle and FTY720 treatment. Right, summary plots of % TCF1 hi (top) and KI67 + (bottom) of EdU + TCR TAG . Each symbol represents an individual mouse, two-way ANOVA followed by post hoc Šídák’s multiple comparisons test. For B-E , n=4-5 mice per group and representative of two independent experiments.

    Article Snippet: TCR TAG transgenic mice (B6.Cg-Tg(TcraY1,TcrbY1)416Tev/J, RRID:IMSR_JAX:005236) , Cre-ER T2 mice (B6.129-Gt(ROSA)26Sortm1(cre/ERT2)Tyj/J, RRID:IMSR_JAX:008463), Rosa26-Cas9 mice (Gt(ROSA)26Sortm1.1(CAG-cas9*,-EGFP)Fezh/J, RRID:IMSR_JAX:026179), B6 Thy1.1 mice (B6.PL-Thy1a/CyJ, RRID:IMSR_JAX:000406), and B6 mice (C57BL/6J, RRID:IMSR_JAX:000664) were purchased from the Jackson Laboratory.

    Techniques: Isolation, Two Tailed Test, Expressing