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