anti mouse tcf 1 c63d9 pe cy7  (Cell Signaling Technology Inc)

Journal: Nature Communications

Article Title: High efficiency CRISPR knock-in demonstrates that TCF1 is insufficient to reverse T cell exhaustion

doi: 10.1038/s41467-026-69671-y

Figure Lengend Snippet: a Schematics of HDR-mediated insertion of TCF1 DNA templates into the endogenous Thy1 or Cx3cr1 genomic loci. Created in BioRender. Chen, A. (2026) https://BioRender.com/t58p429 . ( b – g ) P14 cells were CRISPR edited using the AAV6-Thy1-TCF1 or AAV6-CX3CR1-TCF1 templates and transferred into mice as described in Fig. . Representative ( b , e ) and pooled ( c , d ) data showing frequency ( b ) and fold change (FC) of mean fluorescence intensity (MFI) ( d ) of TCF1 expression in CRISPR edited P14 cells after 8 ( b , c ) or 20 days p.i. ( b , d , e ). Representative ( f ) and pooled ( g ) data showing frequency of Slamf6 + P14 cells after 8 ( g ) or 20 days p.i. f , g Data from 2 independent experiments for day 8 ( n = 8 mice per group for Thy1-TCF1, Thy1 sgRNA and mock groups and n = 7 mice per group for CX3CR1-TCF1 and CX3CR1 sgRNA groups) and data from 3 independent experiments for day 20 ( n = 13 mice for Thy1-TCF1 group and n = 12 mice per group for Thy1 sgRNA, CX3CR1-TCF1, CX3CR1 sgRNA and mock groups). h Pooled data showing TCF1 MFI in edited P14 cells at day 20 p.i. that were transduced with a retrovirus (RV) vector containing TCF1, or CRISPR edited using AAV6-Thy1-TCF1, and transferred into mice as described in Fig. . EV indicates Empty vector. Data from 3 independent experiments ( n = 11 mice per group for RV TCF1 and EV mCherry groups, n = 9 mice for Thy1-TCF1 group and n = 7 mice for mock group). Error bars show ± SEM. p values calculated by ordinary one-way ANOVA with Tukey’s multiple comparison test ( c , g , h and d ).

Article Snippet: For intracellular staining, cells were fixed and permeabilized using eBioscience Foxp3/Transcription factor staining buffer set (ThermoFisher Scientific) and stained with rabbit anti-mouse TCF1 mAb (clone C63D9, Cell Signaling Technology, Cat. no. 2203S, 1:400) detected with a polyclonal secondary anti-rabbit IgG AlexaFluor594 Ab (ThermoFisher Scientific, Cat. no. A-11012, 1:1000), anti-Granzyme-B-APC (Clone GB11, Invitrogen, Cat. no. GRB05, 1:200) and anti-TOX-PE (clone TXRX10, Invitrogen, Cat. no. 12-6502-82, 1:100).

Techniques: CRISPR, Fluorescence, Expressing, Transduction, Plasmid Preparation, Comparison

Journal: Nature Communications

Article Title: High efficiency CRISPR knock-in demonstrates that TCF1 is insufficient to reverse T cell exhaustion

doi: 10.1038/s41467-026-69671-y

Figure Lengend Snippet: a - g P14 cells were CRISPR edited using the AAV6-Thy1-TCF1 or AAV6-CX3CR1-TCF1 template and transferred into mice as described in Fig. . Total P14 cells were sorted 20 days p.i. and analyzed by combined whole cell scRNA-seq and scATAC-seq (multi-ome) analysis. a UMAPs illustrating scRNA-seq, scATAC-seq and Combined data clusters for all cells and for each condition. Proportions of all clusters ( b ) from ( a ), or Tpex cluster only ( c ), per CRISPR genotype in all cells. Heatmaps showing differentially accessible chromatin regions ( d ) and differentially expressed genes ( e ) between the Tpex cluster from Mock, and the CX3CR1 + clusters from each CRISPR genotype. f Gene Set Enrichment Analysis (GSEA) of Tpex or CX3CR1 + gene signatures in CX3CR1 + cells from each TCF1 over-expressing group relative to mock or each other. Fast gene set enrichment analysis (FGSEA) was performed using a two-sided rank-based gene set enrichment test, with gene lists ranked by avg_log2FC as input. Enrichment significance indicated by P values was estimated using a gene permutation-based null distribution with adaptive multilevel Monte Carlo sampling implemented via the fgseaMultiLevel function. P values are adjusted using the Benjamini-Hochberg false discovery rate (FDR) across gene sets within each comparison. NES indicates Normalized Enrichment Score, p adj indicates adjusted p -value ( g ) Heatmap showing transcription factor motif enrichment within differentially accessible chromatin regions in all clusters from each group. Representative and pooled data showing Granzyme-B ( h ), CXCR5 ( i ), PD-1 ( j ) and TOX ( k ) expression in CX3CR1 + CRISPR edited P14 cells at day 20 p.i. gMFI = Geometric mean MFI. Data from 3 independent experiments ( n = 13 mice for Thy1-TCF1 group and n = 12 mice for CX3CR1-TCF1 group). p values calculated by ordinary two-way ANOVA ( h - k ).

Article Snippet: For intracellular staining, cells were fixed and permeabilized using eBioscience Foxp3/Transcription factor staining buffer set (ThermoFisher Scientific) and stained with rabbit anti-mouse TCF1 mAb (clone C63D9, Cell Signaling Technology, Cat. no. 2203S, 1:400) detected with a polyclonal secondary anti-rabbit IgG AlexaFluor594 Ab (ThermoFisher Scientific, Cat. no. A-11012, 1:1000), anti-Granzyme-B-APC (Clone GB11, Invitrogen, Cat. no. GRB05, 1:200) and anti-TOX-PE (clone TXRX10, Invitrogen, Cat. no. 12-6502-82, 1:100).

Techniques: CRISPR, Expressing, Sampling, Comparison

Journal: Cell Reports Medicine

Article Title: Local delivery of IL-15 and anti-PD-L1 nanobody by in vitro- transcribed circILNb elicits superior antitumor immunity in cold tumors

doi: 10.1016/j.xcrm.2025.102413

Figure Lengend Snippet: circILNb reprograms tumor-specific tdLN and tumor CD8 + T cells (A) (Left) Timeline of the experiment designed to evaluate the p-STAT5 levels of CD8 + T cells in tdLN and (right) legends and abbreviations for different treatment groups. B16F10-OVA tumor-bearing mice were intratumorally injected with circIL, circNb, and circILNb (0.5 mg/kg circRNA per mouse). The circLuc was used as control circScram. p value was determined by two-tailed unpaired t test. Data are mean ± SEM. (B) (Left) Representative sample histogram of pSTAT5 after circRNAs treatment. Summary data for (middle) pSTAT5 MFI and (right) pSTAT5 + cells. Gated on live CD3 + CD8 + cells ( n = 3). (C) Timeline of the experiment designed to evaluate the population and activity of CD8 + T cells in tdLN or tumor. B16F10-OVA tumor-bearing mice were intratumorally injected with circIL, circNb, and circILNb (0.5 mg/kg circRNA per mouse). circLuc was used as control circScram. p value was determined by two-tailed unpaired t test. Data are mean ± SEM. (D) (Left) Representative sample histogram of ki67 expression and (right) quantification of the ki67 MFI of tdLN CD8 + T cells. Gated on live CD3 + CD8 + cells ( n = 5). (E) Counts of tetramer + cells in the tdLN, gated on live CD8 + CD44 + tetramer + cells ( n = 5). (F) The tdLN frequency of Ttsm cells, gated on live CD8 + CD44 + tetramer + TCF1 + TOX − cells ( n = 3). (G) (Left) Representative contour plots, (middle) frequency, and (right) counts of Tsle in tumor, gated on live CD8 + tetramer + PD1 + TCF1 + TIM3 - cells ( n = 4). (H) (Left) Frequency and (right) counts of tetramer + cells in the tumor, gated on live CD8 + cells ( n = 3). (I) Counts of IFN-γ + cells in the tumor, gated on live CD8 + tetramer + cells ( n = 5). (J) Counts of GrzmB + cells in the tumor, gated on live CD8 + tetramer + cells ( n = 5). " n " indicates biologically independent samples.

Article Snippet: PE/Cy7 anti-mouse TCF1 (clone: C63D9) , Cell Signaling Technology , Cat#90511S; RRID: AB_3086656.

Techniques: Injection, Control, Two Tailed Test, Activity Assay, Expressing