anti tcf4  (Santa Cruz Biotechnology)

Journal: Cancer Research

Article Title: A Double-Negative Prostate Cancer Subtype Is Vulnerable to SWI/SNF-Targeting Degrader Molecules

doi: 10.1158/0008-5472.CAN-25-2928

Figure Lengend Snippet: TCF7L2 is a dependency in CRPC-WNT. A, Growth data measured by live-cell imaging (IncuCyte S3) upon transfection of indicated siRNA or plasmid (EV or rescue). Immunoblot of indicated proteins at 72 hours after transfection. GAPDH served as loading control. Data are presented as mean values ± SEM and analyzed using two-way ANOVA. ***, P < 0.001. Data are representative of n = 2 independent experiments. B, Spheroid formation measured by live-cell imaging (IncuCyte SX5) upon transfection of indicated siRNA. Data are presented as mean values ± SEM and analyzed using two-way ANOVA. *, P < 0.05; **, P < 0.01; ***, P < 0.001. Data are representative of n = 2 independent experiments. C, IHC and staining intensity of TCF7L2 on indicated organoids upon treatment with 1 µmol/L A858 or 1 µmol/L A947 for 24 hours. Violin plot from TCF7L2 staining intensity analyzed using two-way ANOVA. ****, P < 0.0001. Scale bars, 100 µm (MSK-PCa16) and 50 µm (WCM1078). OD, optical density. D, TOPFlash TCF/LEF reporter assay measured after 48 hours upon treatment with indicated drugs in WCM1078 sublines. FOPFlash served as a negative reporter control. Data are presented as mean values ± SEM after normalization to internal Renilla control and analyzed using a paired Student t test. ***, P < 0.001; ****, P < 0.0001. Data are representative of n = 2 independent experiments.

Article Snippet: The TCF7L2 antibody (Cell Signaling Technology, cat #2569, RRID: AB_2199816) was used for staining.

Techniques: Live Cell Imaging, Transfection, Plasmid Preparation, Western Blot, Control, Staining, Reporter Assay

Journal: Cancer Research

Article Title: A Double-Negative Prostate Cancer Subtype Is Vulnerable to SWI/SNF-Targeting Degrader Molecules

doi: 10.1158/0008-5472.CAN-25-2928

Figure Lengend Snippet: TCF7L2 regulates proproliferative signatures in CRPC-WNT. A, ChIP-seq read density tornado plots from WCM1078 organoids treated with 1 µmol/L A858 or 1 µmol/L A947 for 4 hours ( n = 2 biological replicates). B, Venn diagram indicating A947 treatment–lost regions from ChIP-seq, ATAC-seq, and RNA-seq data in WCM1078. GSEA was performed from 350 overlapping genes. C, Immunoblot of indicated proteins at indicated time upon treatment with 1 µmol/L A947. GAPDH served as loading control. Data are representative of n = 2 independent experiments. D, Dose–response curves with indicated drugs after measurement of proliferation with CellTiter-Glo 2.0 after 7-day treatment ( n = 2 independent experiments).

Article Snippet: The TCF7L2 antibody (Cell Signaling Technology, cat #2569, RRID: AB_2199816) was used for staining.

Techniques: ChIP-sequencing, RNA Sequencing, Western Blot, Control

Journal: bioRxiv

Article Title: Single-cell lung eQTL dataset of Asian never-smokers highlights the roles of alveolar cells in lung cancer etiology

doi: 10.64898/2026.03.26.714500

Figure Lengend Snippet: TCF7L2 is a LUAD susceptibility gene involved in Wnt signaling and promoting cellular growth in the AT2 lineage a, Locus zoom plots of eQTL in AT2 cells for TCF7L2 and GWAS (Shi et al EAS, LUAD) p-values of the region (+/-100 kb) around the GWAS top SNP, rs11169089 in the 10q25.2 locus. LD (r 2 ) relationships between the variants are extracted from 1000G EAS Phase 3 v5. b, Allele frequencies difference between East Asian and European from 1000G c, Association between normalized expression of TCF7L2 and rs1196089 is shown as violin plots. 0 = reference allele, while 1 = alternative; red indicates risk allele, while blue indicates protective. Center line denotes the median, while the 25 th and 75 th percentile is marked as the lower and upper line of the box, respectively. Whiskers extend 1.5 times from the 25 th and 75 th percentiles; outliers are represented as dots. The violin width reflects the density. Note the nominal p-value threshold is 1.31e-05. d, Location of guide RNAs targeting the regions around our SNP(s) of interest. Tukey plot shows GAPDH -normalized mRNA levels of TCF7L2 in A549 cell line from 6 replicates from three independent experiments (n =18). Fold change of target gene expression over non-targeting control is shown as median with interquartile range (IQR) in a box. Whiskers extend 1.5 times IQR, with outliers shown as dots. AAVS1 represents a safe harbor site-targeting gRNA. P-values were calculated using a two-tailed Mann Whitney U test. e and f, GSEA analysis in AT2 cells comparing individuals that highly (75 th percentile) vs lowly (25 th percentile) expressed TCF7L2 . The top 5 most enriched gene sets by normalized enrichment score (NES) are shown. A positive NES indicates enrichment of the gene set compared to the reference. Cumulative enrichment score (e) and NES (f) for the top 5 gene sets are shown with a matching color for each gene set in GSEA (e) and NES (f) plots, where circle sizes are scaled to the-log 10 FDR values. g, A representative xCELLigence growth assay result among 5 independent experiments, where mean cell index with SEM of up to 4 replicates over time is shown. Beta estimates (during growth phase) from linear-mixed-effect models comparing shRNA(s) to control are shown; negative sign is indicative of decreased growth rate compared to control. *** denotes p-value < 1e-04 while ** p-value < 1e-03.

Article Snippet: TCF7L2 knockdown was confirmed by western blotting for TCF7L2 (rabbit monoclonal, Cell Signaling #2569S, 1:1000) and GAPDH (mouse monoclonal, Santa Cruz Biotechnology #47724, 1:500).

Techniques: Expressing, Targeted Gene Expression, Control, Two Tailed Test, MANN-WHITNEY, Growth Assay, shRNA

Journal: bioRxiv

Article Title: A Rare T-Cell Factor 4 Lineage-negative Epithelial Stem Cell Supports Wound Repair and APC-deletion-induced Colon Tumorigenesis

doi: 10.64898/2026.03.17.712502

Figure Lengend Snippet: (A) mGFP marks Tcf4 CreNeo lineage in (Aa,b) full crypts and (Ac,d) differentiated surface epithelium. (B) EdU labeled cells in a Tcf4 Null crypt (mGFP+) isolated from Tcf4 CreNeo/Lox mice are Tcf4 Lin- (mTOM+) cells (yellow arrows). (C) Tcf4 Cre lineage in surface epithelium (Ca,b) and in isolated crypts (Cc-g). Yellow dotted line in (c) represents a region where crypts have been pushed out of the crypt bed. Yellow arrows mark rare Tcf4 Lin- cells that appear at different locations throughout the crypt. (D) Isolation of single crypt cells followed by and FACS analysis (Da) demonstrates the presence of (Db) homogeneous Tcf4 Lin- and heterogeneous (Dc) MIX and (Dd) Tcf4 Lin+ cell populations. (E) Tcf4 Lin- descendants have morphological characteristics of secretory and absorptive cell types (scale bar, (Ea-c) 10 μm: (Ed-e) 20 μm). (F) Model depicting Tcf4 Lineage tracing as stem cells become committed towards absorptive and secretory lineages and (G) the published markers associated with lineage commitment. (H) Co-staining of Tcf4 Lin+ and Lin- cells and commitment markers. Dotted red lines encircle Tcf4 Lin- cells. Green arrows denote Tcf4 Lin- cells that express the commitment marker. Yellow arrows denote Tcf4 Lin- cells that lack commitment marker expression. Red arrows denote Tcf4 Lin+ cells that express the commitment marker.

Article Snippet: All other lines have been backcrossed and maintained in the C57/BL6 background: Tcf4 Cre , Tcf4 CreNeo , Rosa mTmG , Apc LoxEx1-15 (JAX stock #009045; ( )), Apc LoxEx5 (KOMP; APC tm1c; Reporter removed with FlpE recombination , as animals with reporter have a phenotype in the absence of Cre, and no phenotype without reporter until conditional deletion), Cd2 iCre (JAX stock #008520; ( ; )).

Techniques: Labeling, Isolation, Staining, Marker, Expressing

Journal: bioRxiv

Article Title: A Rare T-Cell Factor 4 Lineage-negative Epithelial Stem Cell Supports Wound Repair and APC-deletion-induced Colon Tumorigenesis

doi: 10.64898/2026.03.17.712502

Figure Lengend Snippet: Analysis of single cell RNA-sequencing data utilizing the R package Seurat. (A) tSNE plot clustering Tcf4 Lin- cells into four distinctive groups. (B) Top 10 differentially expressed genes of four clusters of Tcf4 Lin- cells displayed in a heatmap. (C) tSNE plot clustering Epi Tcf4 Lin- cells into three distinctive groups. (D) Top 10 differentially expressed genes of three clusters of Epi Tcf4 Lin- cells displayed in a heatmap. (E) Visualization of several absorptive and secretory precursor and differentiation markers via DotPlot, demonstrating average expression level and the percentage of cells expressing the respective gene in each cluster. (F-I) Feature plots displaying the co-expression of two genes simultaneously on the tSNE plot. (F) Co-expression of Hes1 and Atoh1 . (G) Co-expression of Reg4 and Muc2 . (H) Co-expression of Reg4 and Fcgbp . (I) Co-expression of Reg4 and Clca1 . (J) Visualization of several published stem cell markers via DotPlot, demonstrating average expression level and the percentage of cells expressing the respective gene in each cluster.

Article Snippet: All other lines have been backcrossed and maintained in the C57/BL6 background: Tcf4 Cre , Tcf4 CreNeo , Rosa mTmG , Apc LoxEx1-15 (JAX stock #009045; ( )), Apc LoxEx5 (KOMP; APC tm1c; Reporter removed with FlpE recombination , as animals with reporter have a phenotype in the absence of Cre, and no phenotype without reporter until conditional deletion), Cd2 iCre (JAX stock #008520; ( ; )).

Techniques: Single Cell, RNA Sequencing, Expressing

Journal: bioRxiv

Article Title: A Rare T-Cell Factor 4 Lineage-negative Epithelial Stem Cell Supports Wound Repair and APC-deletion-induced Colon Tumorigenesis

doi: 10.64898/2026.03.17.712502

Figure Lengend Snippet: Analysis of the pseudotime properties of Epi Tcf4 Lin- cells by employing the monocle algorithm. (A) Single cell trajectory analysis, displaying pseudotime, places each cell along the trajectory depending on the amount of transcriptional change each cell has undergone. (B-I) Single cell trajectories displaying the expression level of a marker and placing cells in pseudotime. (B) Sox4 , a distinctive marker of the Stem cell cluster. (C) Muc2 , a distinctive marker of Secretory precursors. (D) Lgals3 , a distinctive marker of Absorptive precursors. (E) Tcf7l2 . (F) Apc . (G) Klf4 . (H) Tcf7l1 . (I) Klf5 . (J-K) Seurat analysis: Feature plots displaying the co-expression of two genes simultaneously on the tSNE plot. (J) Co-expression of Mki67 and Klf4 . (K) Co-expression of Mki67 and Klf5 .

Article Snippet: All other lines have been backcrossed and maintained in the C57/BL6 background: Tcf4 Cre , Tcf4 CreNeo , Rosa mTmG , Apc LoxEx1-15 (JAX stock #009045; ( )), Apc LoxEx5 (KOMP; APC tm1c; Reporter removed with FlpE recombination , as animals with reporter have a phenotype in the absence of Cre, and no phenotype without reporter until conditional deletion), Cd2 iCre (JAX stock #008520; ( ; )).

Techniques: Single Cell, Expressing, Marker

Journal: bioRxiv

Article Title: A Rare T-Cell Factor 4 Lineage-negative Epithelial Stem Cell Supports Wound Repair and APC-deletion-induced Colon Tumorigenesis

doi: 10.64898/2026.03.17.712502

Figure Lengend Snippet: (A-B) Confocal Images, cell number, and crypt location of Epi Tcf4 Lin- and MIX cells (scale bar, 50 μm). Yellow arrows mark unique cellular characteristics (C) EdU labeling and FACs analysis for cell viability (Ca-d) and proliferation (Ce-h) of Epi Tcf4 Lin-, MIX and Tcf4 and Cd2 Lin+ cell populations. (D-E) A representative example of morphological analysis of fluorescence of Epi Tcf4 Lin- and MIX cell populations combined with (D) fluorescent EdU+ and (E) differential interference contrast (Nomarski DIC; scale bar, 10 μm). (F-G) Characterization of Epi Tcf4 Lin- and MIX populations (F) by co-staining with antibodies to known cell markers DCLK1 (Tuft) and PYY (Enteroendocrine L-cells) or (G) by morphology (scale bar in (F) is 20 μm and in (G) is 10 μm). Note that the MIX population is where mGFP and mTOM are labeling membranes in the same cell but are not overlapping.

Article Snippet: All other lines have been backcrossed and maintained in the C57/BL6 background: Tcf4 Cre , Tcf4 CreNeo , Rosa mTmG , Apc LoxEx1-15 (JAX stock #009045; ( )), Apc LoxEx5 (KOMP; APC tm1c; Reporter removed with FlpE recombination , as animals with reporter have a phenotype in the absence of Cre, and no phenotype without reporter until conditional deletion), Cd2 iCre (JAX stock #008520; ( ; )).

Techniques: Labeling, Fluorescence, Staining

Journal: bioRxiv

Article Title: A Rare T-Cell Factor 4 Lineage-negative Epithelial Stem Cell Supports Wound Repair and APC-deletion-induced Colon Tumorigenesis

doi: 10.64898/2026.03.17.712502

Figure Lengend Snippet: Analysis of the wound bed at 3 days (A,C-D) and 5 days (B, E-F) post-endoscopic biopsy. (A-B) Whole mount microscopy of wound bed at 3 days (A) and 5 days (B) post-damage. Yellow lines encircle the wound. (C-D) 3 days following damage, adjacent tissue sections were stained with EPCAM and PYY and (E-F) 5 days following damage sections were stained with EPCAM and EdU, and analyzed along with Tcf4 lineage status. White boxes in Ca,Da,Ea,Fa (scale bar is 200 μm) denote regions of magnification in Cb,Db,Eb,Fb (scale bar is 50 μm). Turquoise arrows mark the WAE, and magenta arrows mark adjacent crypts that initially cover the wound bed. Yellow arrows mark the injury boundary. Red arrows mark PYY positive MIX cells and white arrows mark EdU+ regions of proliferation.

Article Snippet: All other lines have been backcrossed and maintained in the C57/BL6 background: Tcf4 Cre , Tcf4 CreNeo , Rosa mTmG , Apc LoxEx1-15 (JAX stock #009045; ( )), Apc LoxEx5 (KOMP; APC tm1c; Reporter removed with FlpE recombination , as animals with reporter have a phenotype in the absence of Cre, and no phenotype without reporter until conditional deletion), Cd2 iCre (JAX stock #008520; ( ; )).

Techniques: Microscopy, Staining

Journal: bioRxiv

Article Title: A Rare T-Cell Factor 4 Lineage-negative Epithelial Stem Cell Supports Wound Repair and APC-deletion-induced Colon Tumorigenesis

doi: 10.64898/2026.03.17.712502

Figure Lengend Snippet: Structure of the (A) Apc LoxEx1-15 and (B) Apc LoxEx5 alleles. (C) A cartoon depicting the crypt localization patterns of Tcf4 Cre expression, and compared to Hprt Cre Germline patterns (in all cells), and Tcf4 CreNeo in stem cells of rare crypts and differentiated surface epithelium. (D) Genotyping analysis conducted on colon and tail tissue collected from (Da) Apc LoxEx1-15 and (Db) Apc LoxEx5 and negative control mice following Cre-dependent recombination. (E) Survival Curves for Apc Min/+ , Apc Δ Ex1-15 (Hprt Cre Apc LoxEx1-15/+ ) and Apc Δ Ex5 (Hprt Cre Apc LoxEx5/+ ) mice. Median survival days were 159 (n=14), 125 (n=9), and 156 (n=14), respectively. P values by chi-square statistic were calculated using the log rank Mantel-Cox test. (G-H) Whole mount colons (G) and colon tumor counts (H) in colons isolated from 4-month-old Apc LoxEx5 Tcf4 Cre and Apc LoxEx1-15 Tcf4 Cre animals. ***p = 0.0005 using two–sample t- test with Welch correction. Error bars were calculated using SEM. (I) Survival Curves for Tcf4 Cre/+ Apc Min/+ , Tcf4 Cre/+ Apc LoxEx5/+ and Tcf4 Cre/+ Apc LoxEx1-15 with Apc Min/+ animals for comparison. Median survival days were 154 (n=11; Tcf4 Cre/+ Apc Min/+ ) and 136 (n=11; Tcf4 Cre/+ Apc LoxEx5/+ ). At the end of the 8-month study, all Tcf4 Cre/+ Apc LoxEx1-15/+ animals survived and had no visible phenotypes and were censored from the study. P values by chi-square statistic were calculated using the log rank Mantel-Cox test for greater than 3 groups. (J) Histological analysis summary and (K) histology of colon tissue sections of colons isolated from Apc LoxEx1-15 and Apc LoxEx5 conditionally deleted by Hprt Cre Tcf4 Cre and Tcf4 CreNeo driven recombination. (L) Representative endoscopy of the colon in 2 month and 4-month-old Tcf4 Cre/+ Apc LoxEx5/+ animals. (M) colon tumor counts in colons isolated from 4-month-old Apc LoxEx5 Tcf4 CreNeo and Apc LoxEx1-15 Tcf4 CreNeo animals. *p = 0.05 using two–sample t- test with Welch correction. Error bars were calculated using SEM.

Article Snippet: All other lines have been backcrossed and maintained in the C57/BL6 background: Tcf4 Cre , Tcf4 CreNeo , Rosa mTmG , Apc LoxEx1-15 (JAX stock #009045; ( )), Apc LoxEx5 (KOMP; APC tm1c; Reporter removed with FlpE recombination , as animals with reporter have a phenotype in the absence of Cre, and no phenotype without reporter until conditional deletion), Cd2 iCre (JAX stock #008520; ( ; )).

Techniques: Expressing, Negative Control, Isolation, Comparison

Journal: bioRxiv

Article Title: A Rare T-Cell Factor 4 Lineage-negative Epithelial Stem Cell Supports Wound Repair and APC-deletion-induced Colon Tumorigenesis

doi: 10.64898/2026.03.17.712502

Figure Lengend Snippet: Our data suggest a model whereby (A) Epi Tcf4 Lin- cells represent a rare pool of stem cells that migrate to and repair sites of damage by replenishing the epithelium and mucus layer with a 1:1 ratio of absorptive to secretory cells, (B) Epi Tcf4 Lin- stem cells form all cell types of the large intestine upon expression of Tcf4 , and (C) The Epi Tcf4 Lin- cell represents the cell-of-origin for colon tumors driven by heterozygous germline deletion of Apc .

Article Snippet: All other lines have been backcrossed and maintained in the C57/BL6 background: Tcf4 Cre , Tcf4 CreNeo , Rosa mTmG , Apc LoxEx1-15 (JAX stock #009045; ( )), Apc LoxEx5 (KOMP; APC tm1c; Reporter removed with FlpE recombination , as animals with reporter have a phenotype in the absence of Cre, and no phenotype without reporter until conditional deletion), Cd2 iCre (JAX stock #008520; ( ; )).

Techniques: Expressing