Journal: bioRxiv

Article Title: ACB1801 enhances tumor immunogenicity by targeting glycolysis/ferroptosis vulnerability and activating STAT1-signaling to overcome anti-PD-1 resistance in MSS colorectal cancer

doi: 10.64898/2026.02.23.707369

Figure Lengend Snippet: (A) KEGG pathway enrichment analysis of CD45-cells isolated from tumors treated with anti-PD-1 alone or with combination anti-PD-1/ACB1801. The dot plot shows the top 30 enriched KEGG pathways ranked by RichFactor, with dot size indicating the number of genes per pathway and color representing the adjusted p-value for significance (blue to red scale). Pathways associated with ACB1801-related functional changes in tumor CD45-cells are highlighted, including those related to metabolic processes. (B) KEGG pathway annotation of metabolic processes in CD45-cells from tumors treated with anti-PD-1 alone or in combination with ACB1801. The bar graph illustrates the number of genes involved in key metabolic pathways, grouped by category. The greatest gene representation is found in global and overview maps, with additional subdivisions including carbohydrate, lipid, glycan, amino acid, nucleotide, and energy metabolism, as well as biosynthetic and biodegradation pathways. This profile highlights metabolic processes most affected by ACB1801 treatment in the combination setting. (C) Gene set enrichment analysis (GSEA) comparing metabolic pathway activity between ACB1801- and DMSO-treated CT26 cells. The enrichment plots show normalized enrichment scores (NES) and p-values for glycolysis, fatty acid metabolism, and oxidation pathways. In all cases, metabolic pathway gene sets are significantly downregulated in ACB1801-treated cells relative to the DMSO control (p < 0.0001), indicating inhibition of glycolytic and oxidative metabolic programs by ACB1801. (D) Common glycolysis signatures in ACB1801-treated CT26 and HT29 cells. The heatmap displays log2 fold change (Log2FC) for key glycolytic genes in CT26 and HT29 cells following ACB1801 treatment. Each column represents a different gene implicated in glycolysis, while rows correspond to the 2 cell lines. Color scale indicates the degree of gene expression change, highlighting consistent downregulation of glycolytic pathways across both cell types in response to ACB1801. (E) ACB1801 suppresses glycolytic gene expression in CT26 and HT29 cells. The bar graphs show mRNA fold changes of key glycolytic genes in CT26 (top) and HT29 (bottom) cells treated with DMSO or ACB1801 (10 μM for CT26; 20 μM for HT29, 24 hours), as measured by quantitative RT-PCR. ACB1801 consistently downregulates multiple glycolytic genes, including HK2, LDHA, SLC16A1, PDK1, GLUT1, ENO1, and PFKL, in both cell lines. Data are presented as the fold change (FC) relative to the control (DMSO), with mean ± SEM from 3 independent experiments. Statistical significance was determined by an unpaired t-test (*p < 0.05, **p < 0.01, ***p < 0.001). (F) ACB1801 reduces intracellular glucose and lactate levels, as well as extracellular lactate secretion, in CT26 and HT29 cells. The bar graphs show the quantification of intracellular glucose, intracellular lactate, and extracellular lactate in cells treated with DMSO or ACB1801 (10 μM for CT26; 20 μM for HT29, 24hours). Both CT26 and HT29 cells exhibit significant decreases in intracellular glucose and lactate, along with reduced extracellular lactate release following ACB1801 treatment. Data represent the mean ± SEM from 3 independent experiments. Statistical significance was determined by an unpaired t-test (*p < 0.05; **p < 0.01).

Article Snippet: CT26, MC38 and HT29 cell lines were obtained from American Type Culture Collection (ATCC).

Techniques: Isolation, Functional Assay, Glycoproteomics, Activity Assay, Control, Inhibition, Gene Expression, Quantitative RT-PCR

Journal: bioRxiv

Article Title: ACB1801 enhances tumor immunogenicity by targeting glycolysis/ferroptosis vulnerability and activating STAT1-signaling to overcome anti-PD-1 resistance in MSS colorectal cancer

doi: 10.64898/2026.02.23.707369

Figure Lengend Snippet: (A) Expression analysis of ferroptosis suppressor and driver genes in CT26 and HT29 cells following ACB1801 treatment (10 μM for CT26; 20 μM for HT29, 24hours). The heatmap illustrates log2 fold change (Log2FC) in the expression of key genes associated with ferroptosis suppression and induction for CT26 and HT29 cells. The color scale reflects relative up- or downregulation of each gene. Distinct expression patterns highlight the impact of ACB1801 on ferroptosis regulatory networks in both cell lines. (B) ACB1801 downregulates the expression of ferroptosis-related genes in HT29 cells. The bar graphs show mRNA fold changes of key genes involved in ferroptosis regulation following treatment with DMSO or ACB1801 (10 μM for CT26; 20 μM for HT29, 24hours), as measured by quantitative PCR. Significant decreases are observed for several ferroptosis suppressors and antioxidants, including SLC7A11, ATF4, NRF2, GPX4, ENO1, LCN2, GCLC, and GCLM. Data represent the mean ± SEM of the fold change relative to the DMSO-treated controls. Statistical significance was determined by an unpaired t-test (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001). (C) ACB1801 lowers the GSH/GSSG ratio in CT26 and HT29 cells. The bar graphs show the ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG) in cells treated with DMSO or ACB1801 (40 μM and 80 μM). ACB1801 significantly decreases the GSH/GSSG ratio in a dose-dependent manner, indicating increased oxidative stress and reduced cellular antioxidant capacity. Data represent the mean ± SEM from 3 independent experiments. Statistical significance was determined by an unpaired t-test (*p < 0.05). (D) ACB1801 increases oxidized ROS and lipid peroxidation in CT26 and HT29 cells. The flow cytometry histograms show dose-dependent changes in oxidized reactive oxygen species (ROS, top) and lipid peroxidation (bottom) in CT26 and HT29 cells treated with escalating concentrations of ACB1801 compared to DMSO controls. Both cell lines exhibit increased oxidative stress and membrane lipid damage upon 24hours with ACB1801. (E) ACB1801 reduces cell viability via ferroptosis in CT26 and HT29 cells. The bar graphs show cell viability following treatment with DMSO or ACB1801 (20, 40, or 80 μM) in the absence or presence of the ferroptosis inhibitor ferrostatin-1. ACB1801 significantly decreases cell viability in both cell lines, an effect that is reversed by ferrostatin-1 co-treatment, indicating ferroptosis involvement. Data represent the mean ± SEM from 3 independent experiments. Statistical significance was determined by an ordinary one-way ANOVA (*p < 0.05; **p < 0.01; ns = not significant).

Article Snippet: CT26, MC38 and HT29 cell lines were obtained from American Type Culture Collection (ATCC).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Flow Cytometry, Membrane

Journal: bioRxiv

Article Title: ACB1801 enhances tumor immunogenicity by targeting glycolysis/ferroptosis vulnerability and activating STAT1-signaling to overcome anti-PD-1 resistance in MSS colorectal cancer

doi: 10.64898/2026.02.23.707369

Figure Lengend Snippet: (A) Cytokine profiling of CT26 cell culture supernatants after treatment with DMSO or ACB1801 (10 μM) for 24 hours. The left panel shows an increase in Tap1 expression under these conditions. Each duplicate spot represents the relative abundance of an individual cytokine or chemokine. Highlighted boxes mark CXCL10 (1, red), LCN2 (2, green), and CXCL1 (3, green). The adjacent bar graphs show the integrated density quantification for CXCL10, LCN2, and CXCL1. ACB1801 treatment enhances CXCL10 secretion while reducing LCN2 and CXCL1 levels relative to the DMSO control. (B and C) Quantification of CXCL10 expression at the protein (B) and mRNA (C) levels following treatment with DMSO or ACB1801. (B) ELISA measurement of CXCL10 protein secretion in CT26 and HT29 cells treated with ACB1801 (10 μM for CT26; 20 μM for HT29) for 24 hours shows a significant increase in CXCL10 levels compared with DMSO. (C) Quantitative RT-PCR analysis of Cxcl10 (CT26) and CXCL10 (HT29) mRNA expression reveals elevated transcript levels in ACB1801-treated samples. Data represent the mean ± SEM from 3 independent experiments. Statistical significance was determined by an unpaired t-test (**p < 0.01, ***p < 0.001). (D) Quantification of CXCL10 levels in tumor plasma from CT26 tumors following the indicated treatments. CXCL10 concentrations were measured by ELISA. Data represent the mean ± SEM from 3 tumors per treatment group. Statistical significance was determined by an ordinary one-way ANOVA (*p < 0.05). (E) Gene set enrichment analysis (GSEA) of CT26 cells treated with ACB1801 versus DMSO control. ACB1801 treatment significantly enriches the interferon-gamma response gene signature, as reflected by a high normalized enrichment score (NES) and strong statistical significance (P < 0.0001). (F) Western blot analysis of STAT1 phosphorylation at Tyr701 (Y701) in CT26 and HT29 cells following 24 h treatment with DMSO or ACB1801 (10 µM for CT26; 20 µM for HT29). Whole-cell lysates prepared in RIPA buffer were immunoblotted for phospho-STAT1 (Y701), total STAT1, and β-actin as a loading control. Right panels show the ratio of phospho-STAT1 (Y701) to total STAT1, expressed as the mean ± SEM of three independent experiments. Statistical significance was assessed using an unpaired two-tailed t-test with Welch’s correction (p < 0.05). (G) Quantitative RT-PCR analysis of Cxcl10/CXCL10 mRNA expression in CT26 and HT29 cells after 24-hour treatment with DMSO or ACB1801 (10 μM for CT26; 20 μM for HT29), following transfection with control siRNA (siCTRL) or STAT1-targeting siRNAs (CT26: siStat11#9 and #12; HT29: siSTAT1#7 and #8). ACB1801-induced upregulation of Cxcl10/CXCL10 mRNA is attenuated upon STAT1 knockdown. Data are presented as the fold change (FC) relative to the control, with mean ± SEM from three independent experiments. Statistical significance was determined by an ordinary one-way ANOVA (*p < 0.05; **p < 0.01).

Article Snippet: CT26, MC38 and HT29 cell lines were obtained from American Type Culture Collection (ATCC).

Techniques: Cell Culture, Expressing, Control, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Clinical Proteomics, Western Blot, Phospho-proteomics, Two Tailed Test, Transfection, Knockdown

Journal: bioRxiv

Article Title: ACB1801 enhances tumor immunogenicity by targeting glycolysis/ferroptosis vulnerability and activating STAT1-signaling to overcome anti-PD-1 resistance in MSS colorectal cancer

doi: 10.64898/2026.02.23.707369

Figure Lengend Snippet: (A) Depletion of STAT1 with siRNAs targeting mouse Stat1 or human STAT1 . CT26 cells were transfected with siCTRL or 2 siRNAs targeting mouse Stat1 (#7 and #8), and HT29 cells were transfected with siCTRL or 2 siRNAs targeting human STAT1 (#7 and #8). Following treatment with DMSO or ACB1801, Stat1/STAT1 mRNA levels were quantified by RT-PCR. Data represent the mean ± SEM of the fold change (FC) relative to the siCTRL DMSO from 3 independent experiments. Statistical significance was determined by an ordinary one-way ANOVA (ns, not significant; **p < 0.01; ****p < 0.0001). (B) Volcano plot showing differentially expressed genes in CT26 tumors treated with ACB1801 compared to DMSO. Each point represents an individual gene, with log2 fold change (logFC) on the x-axis and −log10(p-value) on the y-axis. Upregulated genes in ACB1801-treated tumors are shown in red and downregulated genes in blue; no significant genes are shown in gray. Dashed vertical lines indicate log2FC cutoffs (±1), and the dashed horizontal line marks the p-value significance threshold.

Article Snippet: CT26, MC38 and HT29 cell lines were obtained from American Type Culture Collection (ATCC).

Techniques: Transfection, Reverse Transcription Polymerase Chain Reaction

Journal: bioRxiv

Article Title: ACB1801 enhances tumor immunogenicity by targeting glycolysis/ferroptosis vulnerability and activating STAT1-signaling to overcome anti-PD-1 resistance in MSS colorectal cancer

doi: 10.64898/2026.02.23.707369

Figure Lengend Snippet: (A) Gene ontology (GO) enrichment network analysis of differentially expressed genes in CT26 tumors treated with ACB1801. Network nodes correspond to enriched GO biological processes, with node size proportional to the number of associated genes and edges indicating functional relationships between GO terms. Adjusted p-values are color-coded to reflect enrichment strength. Major functional clusters center on protein modification, ubiquitin-dependent protein catabolism, and protein processing (highlighted in blue), consistent with coordinated regulation of pathways involved in antigen processing and presentation. (B) Gene set enrichment analysis (GSEA) comparing CT26 tumors treated with ACB1801 versus DMSO reveals significant enrichment of the antigen processing and presentation pathway. The enrichment plot shows a high normalized enrichment score (NES = 1.90) and strong statistical significance (p = 0.000181), indicating robust upregulation of antigen-processing genes following ACB1801 treatment. (C) ACB1801 increases cell-surface MHC-I expression in CT26 and HT29 cells. The representative flow cytometry histograms show H-2Kd (CT26) and HLA-A, B, and C (HT29) expression after treatment with DMSO or ACB1801 (20 μM, 40 μM) for 24 hours. The bar graphs represent the mean fluorescence intensity (MFI) from 3 independent experiments ± SEM. Statistical significance was determined by an unpaired t-test: *p < 0.05, **p < 0.01. (D) ACB1801 enhances the presentation of the OVA_257–264 (SIINFEKL) antigen in MC38 cells. Cells were pulsed with SIINFEKL peptide for 8 hours, followed by treatment with either DMSO or ACB1801 for 24 hours. Surface OVA–H-2Kb complexes were quantified by flow cytometry using PE–OVA–H-2Kb staining. The representative flow cytometry histograms (top) and bar graphs (bottom) show the mean fluorescence intensity (MFI) from 3 independent experiments ± SEM. Statistical significance was determined by an unpaired t-test (*p < 0.05; ****p < 0.0001), demonstrating that ACB1801 significantly increases SIINFEKL presentation. (E) ACB1801 enhances OT-I CD8+ T cell-mediated killing of MC38-OVA tumor cells. MC38-OVA cells were pulsed with SIINFEKL peptide for 8 hours and co-cultured with activated OT-I CD8+ T cells at the indicated effector: target (E:T) ratios in the presence of DMSO or ACB1801. After 48 hours, tumor cell numbers were quantified using Precision Count Beads. The bar graphs show MC38-OVA cell viability (% ± SEM) from 3 independent experiments. Statistical significance was determined by unpaired t-test (ns = not significant; *p < 0.05; ***p < 0.001), demonstrating that ACB1801 significantly enhances OT-I-mediated cytotoxicity, particularly at higher E:T ratios. (F) Quantitative RT-PCR analysis of NLRC5 mRNA levels in CT26 and HT29 cells after treatment with ACB1801 (20 μM) or DMSO for 24 hours. Data are presented as mean fold change (FC) relative to DMSO-treated samples from 3 independent experiments (± SEM). Statistical significance was determined by an unpaired t-test (*p < 0.05; ****p < 0.0001). (G) Flow cytometry analysis of cell surface MHC-I expression in CT26 and HT29 cells following 24-hour treatment with ACB1801 (20 μM) or DMSO. Cells were transfected with either control siRNA (siCtrl) or NLRC5-targeted siRNAs (siNLRC5#17 and #18 for CT26; siNLRC5#11 and #12 for HT29). Representative flow cytometry histograms show H-2KD (CT26, APC conjugated) and HLA-A, B, C (HT29, PE conjugated) staining. The bar graphs show mean fluorescence intensity (MFI), presented as mean ± SEM from 3 independent experiments. Statistical significance was calculated using an ordinary one-way ANOVA : **p < 0.01, *p < 0.05. (H) Quantitative RT–PCR analysis of NLRC5 expression in CT26 and HT29 cells following STAT1 knockdown and ACB1801 treatment. Cells were transfected with either control siRNA (siCtrl) or STAT1-targeting siRNAs (CT26: siSTAT1#9 and #12; HT29: siSTAT1#7 and #8), then treated for 24 hours with DMSO or ACB1801. NLRC5 mRNA levels are presented as the fold change (FC) relative to the control. Data represent the mean ± SEM from 3 independent experiments. Statistical significance was determined by an ordinary one-way ANOVA (*p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: CT26, MC38 and HT29 cell lines were obtained from American Type Culture Collection (ATCC).

Techniques: Functional Assay, Modification, Ubiquitin Proteomics, Expressing, Flow Cytometry, Fluorescence, Staining, Cell Culture, Quantitative RT-PCR, Transfection, Control, Knockdown

Journal: bioRxiv

Article Title: ACB1801 enhances tumor immunogenicity by targeting glycolysis/ferroptosis vulnerability and activating STAT1-signaling to overcome anti-PD-1 resistance in MSS colorectal cancer

doi: 10.64898/2026.02.23.707369

Figure Lengend Snippet: (A) ACB1801 enhances the expression of MHC-I–associated genes in murine (CT26) and human (HT29) cell lines. Cells were treated with ACB1801 (10 μM) or DMSO for 24 hours, and mRNA levels of Tap1/TAP1, Tap2/TAP2, and B2m/B2M were quantified by RT-PCR and normalized to the control. The bar graphs represent the mean fold change from 3 independent experiments ± SEM. Statistical significance was determined by an unpaired t-test: *p < 0.05, **p < 0.01, ****p < 0.0001. (B) Depletion of NLRC5 blocks the ACB1801-mediated induction of MHC-I–related genes. CT26 cells were transfected with control siRNA (siCTRL) or two siRNAs targeting mouse Nlrc5 (siNlrc5 #11 or #18), and HT29 cells were transfected with siCTRL or two siRNAs targeting human NLRC5 (siNLRC5 #11 or #12). Following transfection, cells were treated with DMSO or ACB1801, and mRNA levels of Nlrc5/NLRC5, Tap1/TAP1, and Tap2/TAP2 were measured by quantitative RT-PCR. Data represent the mean ± SEM of the fold change (FC) relative to the siCTRL DMSO from 3 independent experiments. Statistical significance was determined by ordinary one-way ANOVA (*p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001).

Article Snippet: CT26, MC38 and HT29 cell lines were obtained from American Type Culture Collection (ATCC).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control, Transfection, Quantitative RT-PCR

Journal: bioRxiv

Article Title: ACB1801 enhances tumor immunogenicity by targeting glycolysis/ferroptosis vulnerability and activating STAT1-signaling to overcome anti-PD-1 resistance in MSS colorectal cancer

doi: 10.64898/2026.02.23.707369

Figure Lengend Snippet: (A) ACB1801 reduces the secretion of LCN2 in CT26 and HT29 cells. The bar graphs show LCN2 protein levels in the supernatants of CT26 cells treated with DMSO or 10⍰μM ACB1801, and HT29 cells treated with DMSO or 20⍰μM ACB1801. ACB1801 treatment significantly decreases LCN2 secretion in both cell lines. Data are presented as mean ± SEM from 3 independent experiments. Statistical significance was determined by an unpaired t-test (*p < 0.05, **p < 0.01). (B) Association of glycolysis and ferroptosis signatures with T cell subsets in TCGA colorectal cancer (CRC) patients. The scatter plots show correlations between glycolysis signature scores or ferroptosis suppressor scores and the estimated fractions of CD8 T cells or regulatory T cells (Tregs) in CRC tumors from TCGA. A negative correlation is observed between glycolysis or ferroptosis suppressor signatures and CD8 T cell marker, whereas a positive correlation is seen with Treg marker, as indicated by the Pearson correlation coefficients (R) and p-values shown on each plot.

Article Snippet: CT26, MC38 and HT29 cell lines were obtained from American Type Culture Collection (ATCC).

Techniques: Marker

Journal: bioRxiv

Article Title: Molidustat Targets a Synthetic Lethal Vulnerability in APC-Mutant Colorectal Cancer through GSTP1 and PHD2 Co-Inhibition

doi: 10.64898/2026.01.31.702998

Figure Lengend Snippet: A. Percentage of Cleaved Caspase 3/7 positive HT29 (top), and RKO (bottom) cells. Cells were treated with Molidustat for 48 hours at indicated concentrations, 10uM Staurosporine was used as a positive control (100% cell death). Mean + SEM is assessed by unpaired two tailed Student’s t-test, **p<0.01, (ns) non-significant. B. Representative images of Cleaved Caspase-3/7 signal in DMSO, Molidustat (90 μM), and Staurosporine treated cells. Scale bar: 300 μm. C. Representative Western Blot of PHD2 levels in HT29 cells. D. Percentage confluency of HT29 cells post-transfection with the indicated guide RNAs. E. Cleaved Caspase-3/7 signal in HT29 cells post-transfection with the indicated crRNAs. Mean + SEM is assessed by two-way ANOVA, *p<0.05. N = 3 biologically independent experiments.

Article Snippet: Human colorectal cancer cell lines HT29 and RKO were obtained from the American Type Culture Collection (ATCC) and maintained in Dulbecco’s Modified Eagle Medium (DMEM; Sigma-Aldrich, D6429) supplemented with 10% (v/v) fetal bovine serum (FBS; Gibco, 16000044), 1% (v/v) penicillin-streptomycin (Gibco, 15140122), and 2 mM L-glutamine (Sigma-Aldrich, G7513).

Techniques: Positive Control, Two Tailed Test, Western Blot, Transfection

Journal: iScience

Article Title: Long non-coding RNA UCA1 modulates SMARCA2-containing SWI/SNF chromatin remodeling complexes in human colorectal cancer

doi: 10.1016/j.isci.2025.114283

Figure Lengend Snippet: Loss of UCA1 expression induces a stem cell-like phenotype (A) Ability of HT29-derived cellular clones (control ICP, dUCA1E1, and dUCA1E2) to form spheres measured by ELDA. ( n = 5, ELDA estimated stem cell frequencies with 95% confidence intervals, ∗ p < 0.05, ∗∗ p < 0.01). (B) Representative images of non-adherent multicellular culture for 7 days (Microscope Leica DMi8). (C) Size of spheres measured after individual culture of ICP, dUCA1E1, and dUCA1E2 cells in non-adherent conditions ( n = 7, mean ± SD, one-way ANOVA, ∗ p < 0.05). (D) Representative PGC images of non-adherent individual sphere culture for 11 days (Microscope Zeiss Cell discoverer 7). (E) Percentage of different cell populations in HT29, control ICP, dUCA1E1, and dUCA1E2 cells ( n = 3–9, mean ± SD). Cells were labeled for expression of stem cell markers CD44, CD133, and CD166 and sorted by FACS into a pool of cells expressing no markers (−), one of three (+), two of three (++), or all markers (+++). (F) Different pools of cells expressing CSC markers were analyzed for their ability to form spheres by ELDA ( n = 3, ELDA estimated stem cell frequencies with 95% confidence intervals, ∗∗ p < 0.01, ∗∗∗ p < 0.001).

Article Snippet: The colorectal adenocarcinoma cell lines HT29 (HTB-38, RRID:CVCL_0320) and SW480 (RRID:CVCL _0546) were authenticated through the Short Tandem Repeat (STR) DNA profile analysis according to the procedures recommended by the ATCC Institute and the derived cell models tested negative pour mycoplasma.

Techniques: Expressing, Derivative Assay, Clone Assay, Control, Microscopy, Labeling

Journal: iScience

Article Title: Long non-coding RNA UCA1 modulates SMARCA2-containing SWI/SNF chromatin remodeling complexes in human colorectal cancer

doi: 10.1016/j.isci.2025.114283

Figure Lengend Snippet: UCA1 physically interacts with BRM and BRG1 in HT29 cells (A and B) Representative confocal microscope images of PLA assays of HT29 cells with or without 2 μM 5-FU for 24h showing nuclear staining (DAPI, blue), actin (phallodin, green), and dotted interaction signals (PLA, red) of UCA1 with BRM (A) and with BRG1(B). Scale bars indicate 20 μm. Dotted white lines mark enlarged sections with scale bars of 10 μm. (C and D) Percentages of cells with PLA interactions observed in the nuclei (red; 1 spot/nucleus, dark red; multiple spots/nucleus), and in the cytoplasm (green) normalized to negative controls (mean ± SEM, n = 4, one-way ANOVA ∗ p < 0.05). (E) UCA1 transcript enrichment in RNA-IP analysis of HT29 cells with or without 2 μM 5-FU for 24 h; input (5% of total), control IgG (defined as 100%), and IP with either BRM or BRG1 antibodies ( n = 3, mean ± SEM).

Article Snippet: The colorectal adenocarcinoma cell lines HT29 (HTB-38, RRID:CVCL_0320) and SW480 (RRID:CVCL _0546) were authenticated through the Short Tandem Repeat (STR) DNA profile analysis according to the procedures recommended by the ATCC Institute and the derived cell models tested negative pour mycoplasma.

Techniques: Microscopy, Staining, Control

Journal: iScience

Article Title: Long non-coding RNA UCA1 modulates SMARCA2-containing SWI/SNF chromatin remodeling complexes in human colorectal cancer

doi: 10.1016/j.isci.2025.114283

Figure Lengend Snippet: Loss of UCA1 expression induces a stem cell-like phenotype (A) Ability of HT29-derived cellular clones (control ICP, dUCA1E1, and dUCA1E2) to form spheres measured by ELDA. ( n = 5, ELDA estimated stem cell frequencies with 95% confidence intervals, ∗ p < 0.05, ∗∗ p < 0.01). (B) Representative images of non-adherent multicellular culture for 7 days (Microscope Leica DMi8). (C) Size of spheres measured after individual culture of ICP, dUCA1E1, and dUCA1E2 cells in non-adherent conditions ( n = 7, mean ± SD, one-way ANOVA, ∗ p < 0.05). (D) Representative PGC images of non-adherent individual sphere culture for 11 days (Microscope Zeiss Cell discoverer 7). (E) Percentage of different cell populations in HT29, control ICP, dUCA1E1, and dUCA1E2 cells ( n = 3–9, mean ± SD). Cells were labeled for expression of stem cell markers CD44, CD133, and CD166 and sorted by FACS into a pool of cells expressing no markers (−), one of three (+), two of three (++), or all markers (+++). (F) Different pools of cells expressing CSC markers were analyzed for their ability to form spheres by ELDA ( n = 3, ELDA estimated stem cell frequencies with 95% confidence intervals, ∗∗ p < 0.01, ∗∗∗ p < 0.001).

Article Snippet: colorectal adenocarcinoma cell line HT29 , ATCC , HTB-38 RRID:CVCL_0320.

Techniques: Expressing, Derivative Assay, Clone Assay, Control, Microscopy, Labeling

Journal: iScience

Article Title: Long non-coding RNA UCA1 modulates SMARCA2-containing SWI/SNF chromatin remodeling complexes in human colorectal cancer

doi: 10.1016/j.isci.2025.114283

Figure Lengend Snippet: UCA1 physically interacts with BRM and BRG1 in HT29 cells (A and B) Representative confocal microscope images of PLA assays of HT29 cells with or without 2 μM 5-FU for 24h showing nuclear staining (DAPI, blue), actin (phallodin, green), and dotted interaction signals (PLA, red) of UCA1 with BRM (A) and with BRG1(B). Scale bars indicate 20 μm. Dotted white lines mark enlarged sections with scale bars of 10 μm. (C and D) Percentages of cells with PLA interactions observed in the nuclei (red; 1 spot/nucleus, dark red; multiple spots/nucleus), and in the cytoplasm (green) normalized to negative controls (mean ± SEM, n = 4, one-way ANOVA ∗ p < 0.05). (E) UCA1 transcript enrichment in RNA-IP analysis of HT29 cells with or without 2 μM 5-FU for 24 h; input (5% of total), control IgG (defined as 100%), and IP with either BRM or BRG1 antibodies ( n = 3, mean ± SEM).

Article Snippet: colorectal adenocarcinoma cell line HT29 , ATCC , HTB-38 RRID:CVCL_0320.

Techniques: Microscopy, Staining, Control

Journal: BMC Biology

Article Title: Increased prolactin levels in pregnancy affect colorectal cancer aggressiveness

doi: 10.1186/s12915-025-02500-8

Figure Lengend Snippet: Prolactin sensitizes HT29 cells to TRAIL-mediated apoptosis. A Representative flow cytometry plots of the HT29 cells treated with 0.1 to 1000 ng/mL of soluble TRAIL in static conditions, under FSS, after PRL pre-exposure, and in FSS combined with PRL pre-exposure. B Quantification of the cell viability after soluble TRAIL treatment (statistical analysis was performed comparing all groups to the static 0 ng/mL TRAIL dosage without PRL or FSS). C Quantification of the TRAIL sensitization by each of the experimental groups (statistical analysis was performed comparing all groups to the 0.1 ng/mL TRAIL group without PRL or FSS). D Representative flow cytometry plot and quantification of cell viability, apoptosis, and necrosis of the HT29 cells treated with the DA liposomes in whole blood under FSS conditions. E Representative flow cytometry plot and quantification of cell viability, apoptosis, and necrosis of the PRL pre-exposed HT29 cells treated with the DA liposomes in whole blood under FSS conditions. Statistical significance was evaluated by either an ordinary one-way ANOVA or a two-way ANOVA and is shown as * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 ( n = 3 biological replicates)

Article Snippet: The human colon adenocarcinoma cell lines HT29 (HTB38, ATCC) and COLO320DM (CCL-220, ATCC) were used throughout the study.

Techniques: Flow Cytometry, Liposomes