dld1 cells  (ATCC)

Journal: Frontiers in Pharmacology

Article Title: Integrated network pharmacology and experimental verification to reveal the mechanisms of curcumin in the treatment of colorectal cancer

doi: 10.3389/fphar.2025.1703562

Figure Lengend Snippet: Venn diagram of curcumin targets, CRC-related genes, and DEGs from curcumin-treated DLD1 cells.

Article Snippet: The human CRC cell line DLD1 (ATCC, United States) was cultured as described and seeded in 96-well plates to reach 80% confluence before treatment.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Integrated network pharmacology and experimental verification to reveal the mechanisms of curcumin in the treatment of colorectal cancer

doi: 10.3389/fphar.2025.1703562

Figure Lengend Snippet: Curcumin promotes ferroptosis and regulates Wnt/β-catenin signaling pathway in vitro . (A) Representative images of transmission electron microscopy observation of mitochondria; curcumin induced reduced mitochondrial volume and increased membrane density in HCT116 cell lines (scale bar: 2 μm and 500 nm); (B) Relative mitochondrial area in HCT116 cell lines. (C) Relative mitochondrial density in HCT116 cell lines. (D–E) Curcumin induced the accumulation of Lipid ROS in CRC cells. Dose-dependent accumulation of Lipid ROS in HCT116 and DLD1 cells treated with curcumin (5–20 μm). (F–G) Curcumin effectively inhibited the expression of SLC7A11 and GPX4, downregulated level of β-catenin and promoted phosphorylation of GSK3β at Ser9. *P < 0.05, **P < 0.01 and ***P < 0.001.

Article Snippet: The human CRC cell line DLD1 (ATCC, United States) was cultured as described and seeded in 96-well plates to reach 80% confluence before treatment.

Techniques: In Vitro, Transmission Assay, Electron Microscopy, Membrane, Expressing, Phospho-proteomics

Journal: iScience

Article Title: SLC7A11-specific CAR-T cell therapy potently targets colorectal and pancreatic cancer

doi: 10.1016/j.isci.2025.113713

Figure Lengend Snippet: Antibody design and affinity evaluation (A) The procedures for generating murine monoclonal antibody using hybridoma technology. (B) The qPCR experiment confirmed the successful knockdown of SLC7A11 expression levels in DLD1 cells mediated by lentivirus ( n = 3). (C) The specificity of the antibody targeting SLC7A11 was validated using flow cytometry ( n = 4). (D) The affinity between 1A4 and SLC7A11 was confirmed through SPR analysis. (E) The affinity between humanized 1A4 and SLC7A11 was confirmed through SPR analysis. The SLC7A11 expression level data by qPCR were analyzed by t test. The MFI of SLC7A11 data was analyzed by ANOVA test. Data were presented as mean ± SD. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The human colorectal cancer and pancreatic cell lines DLD1 (RRID: CVCL_0248), HT29 (RRID: CVCL_0320), HCT116 (RRID: CVCL_0291), PANC-1 (RRID: CVCL_0480) and MIA PaCa-2 (RRID: CVCL_0428) were obtained from the American Type Culture Collection.

Techniques: Knockdown, Expressing, Flow Cytometry

Journal: iScience

Article Title: SLC7A11-specific CAR-T cell therapy potently targets colorectal and pancreatic cancer

doi: 10.1016/j.isci.2025.113713

Figure Lengend Snippet: The cytotoxicity assays in vitro by CCK8, LDH, and MFI detection (A) The survival rate of target cells after 24 h of co-culture ( n = 5). (B) The lysis rates of target cells after 24 h of co-culture with UTD, MOCK, and CAR-T cells ( n = 3). (C) The co-culture figures under the fluorescence microscope for the Mock group and CART group ( n = 3). Scale bars represent 50 μm. (D andE) The cytotoxic effects of DLD1 cells by measuring the fluorescence intensity after co-culturing ( n = 3). Scale bars represent 50 μm. Data were analyzed by ANOVA test. The CCK8 data and MFI data were presented as mean ± SD. The LDH data were presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: The human colorectal cancer and pancreatic cell lines DLD1 (RRID: CVCL_0248), HT29 (RRID: CVCL_0320), HCT116 (RRID: CVCL_0291), PANC-1 (RRID: CVCL_0480) and MIA PaCa-2 (RRID: CVCL_0428) were obtained from the American Type Culture Collection.

Techniques: In Vitro, Co-Culture Assay, Lysis, Fluorescence, Microscopy

Journal: bioRxiv

Article Title: CIP2A Mediates the Recruitment of the SLX4-MUS81-XPF Tri-Nuclease Complex in Mitosis and Protects Against Replication Stress

doi: 10.1101/2025.09.19.677284

Figure Lengend Snippet: A) Dot plot showing CIP2A foci per mitotic cells in DLD1 WT and DLD1 BRCA2 knockout cells. Statistical analysis: Mann-Whitney test, n = 2 independent replicates. B) Dot plot showing TOPBP1 foci per mitotic cells in DLD1 WT and DLD1 BRCA2 knockout cells. Statistical analysis: Mann-Whitney test, n = 2 independent replicates. C) Quantitative analysis of the mean proximity ligation assay (PLA) intensity in DLD1 WT and DLD1 BRCA2 knockout cells stained for CIP2A and TOPBP1. Statistical analysis: Welch’s t-test. n = 2 independent replicates. D) Dot plot showing CIP2A foci per mitotic cell in U2OS cells treated with ATR inhibitor (ATRi, 10 μM) for 24 h following siRNA depletion. Cells were synchronized in RO-3306 (7 μM) for 6 h. Mean indicated. Statistical analysis: Kruskal-Wallis test, followed by uncorrected Dunn’s multiple comparisons test. n = 2 independent replicates. E) Dot plot showing CIP2A foci per mitotic cell in DLD1 cells treated with Aph (0.4 μM) for 24 h. Cells were synchronized in RO-3306 (7 μM) for 7 h. Median indicated. Statistical analysis: Kruskal-Wallis test, followed by uncorrected Dunn’s multiple comparisons test. n = 4 independent replicates. F) Dot plot showing TOPBP1 foci per mitotic cell in DLD1 cells treated with Aph (0.4 μM) for 24 h. Cells were synchronized in RO-3306 (7 μM) for 7 h. Median indicated. Statistical analysis: Kruskal-Wallis test, followed by uncorrected Dunn’s multiple comparisons test. n = 3 independent replicates. G) Dot plot showing CIP2A foci per mitotic cell in U2OS cells treated with Aph (0.4 μM) for 24 h following siRNA depletion. Cells were synchronized in RO-3306 (7 μM) for 7 h. Median indicated. Statistical analysis: Kruskal-Wallis test, followed by uncorrected Dunn’s multiple comparisons test. n = 3 independent replicates. H) Confocal microscopy images showing CIP2A foci (red) in mitotic U2OS cells, with nuclei stained with DAPI (blue). Images are maximum projections of Z-stacks. I) Dot plot showing TOPBP1 foci per mitotic cell in U2OS cells treated with Aph (0.4 μM) for 24 h following siRNA depletion. Cells were synchronized in RO-3306 (7 μM) for 7 h. Median indicated. Statistical analysis: Kruskal-Wallis test, followed by uncorrected Dunn’s multiple comparisons test. n = 2 independent replicates. J) Confocal microscopy images showing TOPBP1 foci (green) in mitotic U2OS cells, with nuclei stained with DAPI (blue). Images are maximum projections of Z-stacks.

Article Snippet: DLD1 BRCA2 knockout (KO) cells were obtained from ATCC (HTB-96).

Techniques: Knock-Out, MANN-WHITNEY, Proximity Ligation Assay, Staining, Confocal Microscopy

Journal: bioRxiv

Article Title: CIP2A Mediates the Recruitment of the SLX4-MUS81-XPF Tri-Nuclease Complex in Mitosis and Protects Against Replication Stress

doi: 10.1101/2025.09.19.677284

Figure Lengend Snippet: A) Dot plot showing FANCD2 foci per mitotic cell in U2OS cells treated as illustrated in the above panel, with median indicated. n = 3 independent replicates. B) Dot plot showing SLX4 foci per mitotic cell in U2OS cells treated as illustrated in the above panel. Median indicated. n = 2 independent replicates. C) Dot plot showing XPF foci per mitotic cell in U2OS cells treated as illustrated in the above panel. Median indicated. n = 2 independent replicates. D) Confocal microscopy images showing MUS81 foci (red) in mitotic U2OS cells, with nuclei stained with DAPI (blue). Images are maximum projections of Z-stacks. E) Dot plot showing MUS81 foci per mitotic cell in U2OS cells treated as illustrated in the above panel, with median indicated. n = 2 independent replicates. F) Dot plot showing MUS81 foci per mitotic cell in DLD1 cells treated as illustrated in panel depicted in E) , with median indicated. n = 2 independent replicates. G) Dot plot showing MUS81 foci per mitotic cell in DLD1 cells treated as in panel depicted in E) . Mean indicated. n = 4 independent replicates. H) Dot plot showing RAD52 foci per mitotic cell in U2OS cells treated as illustrated in the above panel, with median indicated. n = 3 independent replicates. Statistical analysis in all dot plot panels: Kruskal-Wallis test, followed by uncorrected Dunn’s multiple comparisons test.

Article Snippet: DLD1 BRCA2 knockout (KO) cells were obtained from ATCC (HTB-96).

Techniques: Confocal Microscopy, Staining

Journal: bioRxiv

Article Title: CIP2A Mediates the Recruitment of the SLX4-MUS81-XPF Tri-Nuclease Complex in Mitosis and Protects Against Replication Stress

doi: 10.1101/2025.09.19.677284

Figure Lengend Snippet: A) Dot plot showing EdU foci per chromosome in DLD1 cells, with each dot representing a single metaphase spread. Cells were synchronized with RO-3306 for 4 hours, then released in presence of Colcemid and EdU. Median indicated. n = 2 independent replicates. B) Dot plot showing EdU foci per mitotic cell in DLD1 cells treated as illustrated in the above depicted panel. Mean indicated. n = 2 independent replicates. C) Dot plot showing EdU foci per mitotic cell in U2OS cells treated as illustrated in the above depicted panel. Median indicated. n = 3 independent replicates. D) Confocal microscopy images showing EdU foci (green) in mitotic U2OS cells, with nuclei stained with DAPI (blue). Images are maximum projections of Z-stacks. E) Dot plot showing EdU foci per mitotic cell in DLD1 cells treated as in panel depicted in C) , with median indicated. n = 2 independent replicates. F) Dot plot showing EdU foci per mitotic cell in DLD1 cells including DLD1 CIP2A KO complemented with CIP2A WT, treated as in panel depicted in C) . Mean indicated. n = 2 independent replicates. G) Widefield microscopy images showing EdU foci (yellow) on DLD1 WT and CIP2A knockout metaphase spreads stained with DAPI (blue). White arrows indicate some representative EdU foci. H) Dot plot showing EdU foci per chromosome in DLD1 cells, with each dot representing a single metaphase spread. Cells were treated ± Aph for 24 hours and synchronized with RO-3306 during the last 4 hours, then released in presence of Colcemid and EdU for 40 min. Median indicated. n = 3 independent replicates. I) Quantitative analysis of EdU foci pattern in metaphase spreads shown in G) . EdU foci were classified as single, twin, or triplet. n = 3 indpendent replicates. Statistical analysis in all dot plot panels: Kruskal-Wallis test, followed by uncorrected Dunn’s multiple comparisons test.

Article Snippet: DLD1 BRCA2 knockout (KO) cells were obtained from ATCC (HTB-96).

Techniques: Confocal Microscopy, Staining, Microscopy, Knock-Out

Journal: bioRxiv

Article Title: CIP2A Mediates the Recruitment of the SLX4-MUS81-XPF Tri-Nuclease Complex in Mitosis and Protects Against Replication Stress

doi: 10.1101/2025.09.19.677284

Figure Lengend Snippet: A) Schematic representation of the experimental workflow. B) Quantitative analysis of the mean proximity ligation assay (PLA) intensity in DLD1 WT B6L expressing cells treated as in A) and stained for CIP2A and TOPBP1. Statistical analysis: Brown-Forsythe and Welch ANOVA test. n = 2 independent replicates. C) Dot plot showing CIP2A foci per mitotic cell in DLD1 WT B6L expressing cells treated as in A) , with mean indicated. Cells were incubated with Shield-1 (S1) during release. Statistical analysis: Kruskal-Wallis test, followed by uncorrected Dunn’s multiple comparisons test. n = 2 independent replicates. D) Dot plot showing TOPBP1 foci per mitotic cell in DLD1 WT B6L expressing cells treated as in A) , with mean indicated. Cells were incubated with Shield-1 (S1) during release. Statistical analysis: Kruskal-Wallis test, followed by uncorrected Dunn’s multiple comparisons test. n = 2 independent replicates. E) Confocal microscopy images showing EdU foci (green) in mitotic DLD1 WT B6L expressing cells, with nuclei stained with DAPI (blue). Images are maximum projections of Z-stacks. F) Dot plot showing EdU foci per mitotic cell in DLD1 WT B6L expressing cells treated as in A) . Cells were incubated with EdU and Shield-1 (S1) during release. Mean indicated. Statistical analysis: Kruskal-Wallis test, followed by uncorrected Dunn’s multiple comparisons test. n = 2 independent replicates. G) Confocal microscopy images showing MUS81 foci (red) in mitotic DLD1 WT B6L expressing cells, with nuclei stained with DAPI (blue). Images are maximum projections of Z-stacks. H) Dot plot showing MUS81 foci per mitotic cell in DLD1 WT B6L expressing cells treated as in A) . Cells were incubated with Shield-1 (S1) during release. Mean indicated. Statistical analysis: Kruskal-Wallis test, followed by uncorrected Dunn’s multiple comparisons test. n = 2 independent replicates.

Article Snippet: DLD1 BRCA2 knockout (KO) cells were obtained from ATCC (HTB-96).

Techniques: Proximity Ligation Assay, Expressing, Staining, Incubation, Confocal Microscopy