Journal: iScience

Article Title: Condensin upregulation induces intestinal stem cell senescence, driving cell death and increased intestinal permeability

doi: 10.1016/j.isci.2026.117884

Figure Lengend Snippet: Elevated levels of NCAPD3 in immortalized mammalian IECs induce heterochromatin reorganization (A) mCherry transfected and mCherry-NCAPD3 transfected HT29 cell lysates were immunoblotted for NCAPD3 levels or the loading control, β-tubulin. Endogenous NCAPD3 band intensities were quantified from film and normalized to β-tubulin; quantification is shown below the blot. mCherry-NCAPD3 intensity was also quantified and compared to the endogenous levels of NCAPD3; quantitation is shown below the blot. Image shown is representative of three independent experiments. p value was determined by performing an unpaired Student’s t test. statistical analyses. ∗∗ p ≤ 0.01. Error estimates indicate standard deviations from the mean. (B and C) Immunofluorescence analyses were performed to detect mCherry protein (red) and trimethylated lysine 9 on histone H3 (H3K9me3; green) in (B) mCherry transfected or (C) mCherry-NCAPD3 transfected cells. All images were taken with the 63× objective lens on the confocal microscope. Maximum projections of z stacks are shown. Scale bars, 3 μm. Inset shows enlarged views and dotted circles highlight individual DAPI dense foci. Images shown are representative of images obtained from three independent biological replicates.

Article Snippet: Adenocarcinoma derived cell line from human: HT29 , ATCC , HTB-38.

Techniques: Transfection, Control, Quantitation Assay, Immunofluorescence, Confocal, Microscopy

Journal: iScience

Article Title: Condensin upregulation induces intestinal stem cell senescence, driving cell death and increased intestinal permeability

doi: 10.1016/j.isci.2026.117884

Figure Lengend Snippet: NCAPD3 upregulation increases numbers of senescent, mammalian IECs and induces SASP (A) Senescence-associated-β-galactosidase (SA-β-gal) assays were performed in untransfected, mCherry, and mCherry-NCAPD3 transfected HT29 cells at 48 h, 72 h, and 96 h post transfection. Images were taken with a 20× objective lens. Arrows indicate blue, SA-β-gal+ cells. Scale bars, 75μm. (B) Transfected cells containing SA-β-gal+ nuclei were quantified in three separate experiments for each time point. For each experiment, quantified cells > 100. (C) Experiments described in (A) and (B) were also performed in IEC6 cells. For each experiment shown, quantified cells > 70. In (B) and (C), p values were determined by performing unpaired Student’s t-tests. ∗ p ≤ 0.05, ∗∗∗ p ≤ 0.001, ∗∗∗∗ p ≤ 0.0001, ns, not significant. Error bars indicate standard deviations from the mean. (D) Immunofluorescence analyses were performed on untransfected, mCherry, and mCherry-NCAPD3 transfected HT29 cells to detect p21 at 72 h post-transfection. Images were taken on the confocal microscope, using a 40× objective lens. Maximum projections of z stacks are shown. Scale bars, 25 μm. (E) p21 intensity levels were quantified from three independent experiments in (D), using Volocity software. Quantified cells > 400 for each experiment and averages are shown. p values were determined by performing unpaired Student’s t tests. ∗∗∗∗ p ≤ 0.0001. Error bars indicate standard deviations from the mean. (F) Identical experiments as in (D) and (E) were performed in IEC6 cells. (G) RT-qPCR was performed to detect transcript levels of genes activated as part of the senescence associated secretory phenotype (SASP) in mCherry or mCherry-NCAPD3 transfected HT29 cells. p values were determined by performing unpaired Student’s t tests. ∗ p ≤ 0.05, ∗∗ p ≤ 0.01. Error bars indicate standard deviations from the mean.

Article Snippet: Adenocarcinoma derived cell line from human: HT29 , ATCC , HTB-38.

Techniques: Transfection, Immunofluorescence, Confocal, Microscopy, Software, Quantitative RT-PCR

Journal: iScience

Article Title: Condensin upregulation induces intestinal stem cell senescence, driving cell death and increased intestinal permeability

doi: 10.1016/j.isci.2026.117884

Figure Lengend Snippet: Elevated NCAPD3 expression increases cell death in mammalian IECs (A) Immunofluorescence analyses were performed on mCherry, and mCherry-NCAPD3 transfected HT29 cells (red) to detect cell death marker, cleaved-caspase 3 (green). Images were taken on the confocal microscope, using the 40× objective lens. Scale bars, 25 μm. Maximum projections of z stacks are shown. (B) Quantifications of three separate experiments for 72 h and 96 h post transfection are shown. For each of three independent experiments shown, quantified cells > 100. (C) Experiments described in (A) and (B) were performed using IEC6 cells. For each of three independent experiments shown, quantified cells > 70. In (B) and (C), p values were determined by performing unpaired Student’s t tests. ∗ p ≤ 0.05, ∗∗∗ p ≤ 0.001. Error bars indicate standard deviations from the mean.

Article Snippet: Adenocarcinoma derived cell line from human: HT29 , ATCC , HTB-38.

Techniques: Expressing, Immunofluorescence, Transfection, Marker, Confocal, Microscopy

Journal: iScience

Article Title: Condensin upregulation induces intestinal stem cell senescence, driving cell death and increased intestinal permeability

doi: 10.1016/j.isci.2026.117884

Figure Lengend Snippet:

Article Snippet: Adenocarcinoma derived cell line from human: HT29 , ATCC , HTB-38.

Techniques: Derivative Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lipid Accumulation Promotes Tumorigenic Behaviour of Colorectal Cancer Cells

doi: 10.1111/jcmm.71393

Figure Lengend Snippet: Effect of oleate treatment on cellular lipid accumulation in CRC cells. HT29 and Caco‐2 cells were incubated with indicated oleate (OL) concentrations for 3–48 h. (A) Intracellular triglyceride levels were normalized to cellular protein content. (B) Representative light microscopic images of oil red O staining (400× magnification). (C) Free fatty acids in the supernatant of cells incubated with 0.4 mM OL at indicated time points. (D) PLIN2 mRNA expression analysed by quantitative RT‐PCR and protein expression analysed by western blot. (E) Representative images of immunofluorescence staining for PLIN2 (red) (Magn. 400×). Cell nuclei were stained with DAPI (blue) (* p < 0.05 compared to control (Ctr)).

Article Snippet: The human CRC cell lines HT29 (ATCC HTB‐38), Caco‐2 (ATCC HTB‐37), SW48 (ATCC CCL‐231), SW480 (ATCC CCL‐228), HCT116 (ATCC CCL‐247) and LoVo (ATCC CCL‐229) were cultured in DMEM supplemented with penicillin (400 U/mL), streptomycin (50 μg/mL), L‐glutamine (5840 mg/L), glucose (450 mg/dL) and 10% fetal calf serum (FCS) and incubated at 37°C in an 8% CO 2 atmosphere as described in [ ].

Techniques: Incubation, Staining, Expressing, Quantitative RT-PCR, Western Blot, Immunofluorescence, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lipid Accumulation Promotes Tumorigenic Behaviour of Colorectal Cancer Cells

doi: 10.1111/jcmm.71393

Figure Lengend Snippet: Effect of lipid uptake on expression of key enzymes of lipid metabolism in CRC cells. HT29 and Caco‐2 cells were treated with indicated oleate (OL) concentrations for 12–24 h. (A) ACLY mRNA expression. (B) FASN mRNA expression. (C) SCD1 mRNA expression. (D) DGAT2 mRNA expression and (E) CPT1A mRNA expression analysed by quantitative RT‐PCR and CPT1A protein expression analysed by western blot. (F) ATGL protein expression analysed by western blot (* p < 0.05 compared to control (Ctr)).

Article Snippet: The human CRC cell lines HT29 (ATCC HTB‐38), Caco‐2 (ATCC HTB‐37), SW48 (ATCC CCL‐231), SW480 (ATCC CCL‐228), HCT116 (ATCC CCL‐247) and LoVo (ATCC CCL‐229) were cultured in DMEM supplemented with penicillin (400 U/mL), streptomycin (50 μg/mL), L‐glutamine (5840 mg/L), glucose (450 mg/dL) and 10% fetal calf serum (FCS) and incubated at 37°C in an 8% CO 2 atmosphere as described in [ ].

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Lipid Accumulation Promotes Tumorigenic Behaviour of Colorectal Cancer Cells

doi: 10.1111/jcmm.71393

Figure Lengend Snippet: Effect of cellular lipid accumulation on tumorigenicity of cancer cells. HT29 and Caco‐2 cells were treated with indicated oleate (OL) concentrations for 12–24 h. (A) Proliferation and (B) migratory activity of CRC cells. mRNA expression of EMT‐markers (C) E‐CAD and N‐CAD and (D) SNAIL (E) and TWIST. (F) mRNA expression of IL‐8 (* p < 0.05 compared to control (Ctr)).

Article Snippet: The human CRC cell lines HT29 (ATCC HTB‐38), Caco‐2 (ATCC HTB‐37), SW48 (ATCC CCL‐231), SW480 (ATCC CCL‐228), HCT116 (ATCC CCL‐247) and LoVo (ATCC CCL‐229) were cultured in DMEM supplemented with penicillin (400 U/mL), streptomycin (50 μg/mL), L‐glutamine (5840 mg/L), glucose (450 mg/dL) and 10% fetal calf serum (FCS) and incubated at 37°C in an 8% CO 2 atmosphere as described in [ ].

Techniques: Activity Assay, Expressing, Control

Journal: Virulence

Article Title: Activity of Lactobacillus gasseri strain LLG1 and its exopolysaccharides in an inflammatory cell model and on Candida albicans proliferation

doi: 10.1080/21505594.2026.2707757

Figure Lengend Snippet: Effect of Lg -EPS on the expression of inflammatory receptors or mediators of macrophages, on levels of IL-6 or IL-10 secreted by macrophages exposed to LPS, and induction of AhR activity by Lg -EPS in HT29-Lucia™ AhR cells. (A–H) Relative mRNA expression levels of TLR2 (A), TLR4 (B), Myd88 (C), NF-κB (D), AhR (E), IL-10 (F), IL-6 (G) and TNFα (H), respectively, in macrophages. MΦ represents a control group (macrophages alone); 250, 500 and 1000 correspond to macrophages treated with Lg -EPS at 250, 500 or 1000 µg/mL, respectively. LPS represents a positive control (macrophages exposed to LPS). LPS 250, LPS 500 or LPS 1000 correspond to macrophages exposed to LPS and treated with Lg -EPS at 250, 500 or 1000 µg/mL, respectively. (I-J) Release of IL-6 (I) and IL-10 (J) by macrophages exposed to Lg -EPS at 1 mg/mL. MΦ represents a control group (macrophages alone); Lg -EPS corresponds to macrophages treated with EPS. LPS represents a positive control (macrophages exposed to LPS); LPS + Lg -EPS corresponds to macrophages exposed to LPS and treated with EPS. (K) Induction of AhR activity by Lg -EPS in HT29-Lucia™ AhR cells. HT29 represents a control group (cells alone); FICZ represents a positive control (AhR ligand; 5 µg/mL); CH-223191 represents a negative control (AhR inhibitor; 5 µg/mL); 250, 500 and 1000 correspond to macrophages treated with Lg -EPS at 250, 500 or 1000 µg/mL, respectively. Data represent three independent biological replicates per condition. Relative mRNA expression levels and cytokine release by macrophages (A-J) were assessed in triplicate, whereas induction of AhR activity in HT29-Lucia TM AhR cells (K) was assessed in sextuplicate.

Article Snippet: HT29-Lucia TM AhR cells were also used (AhR reporter cells; ht2l-ahr; InvivoGen), in DMEM containing 10% FBS, 100 U/ml-100 μg/mL of penicillin-streptomycin, 100 μg/mL of Normocin TM (ant-nr-05; InvivoGen) and 100 μg/ml of Zeocin TM (selective antibiotics; ant-zn-1; InvivoGen).

Techniques: Expressing, Activity Assay, Control, Positive Control, Negative Control