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adenocarcinoma derived cell line from human ht29 (ATCC)

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ATCC adenocarcinoma derived cell line from human ht29

Elevated levels of NCAPD3 in immortalized mammalian IECs induce heterochromatin reorganization (A) mCherry transfected and mCherry-NCAPD3 transfected <t>HT29</t> 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.

Adenocarcinoma Derived Cell Line From Human Ht29, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 14265 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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adenocarcinoma derived cell line from human ht29 - by Bioz Stars, 2026-11

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1) Product Images from "Condensin upregulation induces intestinal stem cell senescence, driving cell death and increased intestinal permeability"

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

Journal: iScience

doi: 10.1016/j.isci.2026.117884

Figure Legend 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.

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

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.

Figure Legend 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.

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

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.

Figure Legend 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.

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

Figure Legend Snippet:

Techniques Used: Derivative Assay

Cell Culture:

Article Title: Liposomal encapsulation of cholecalciferol mitigates in vivo toxicity and delays tumor growth
Article Snippet: .. Adherent murine MC38 (University of Washington, Seattle, USA) and human HT29 (ATCC; HTB-38) colorectal cancer cells were cultured with RPMI 1640 (Gibco) supplemented with 10% (v/v) fetal bovine serum (FBS; Gibco), 100 IU penicillin and 100 μg/mL streptomycin (1% P/S; Gibco). ..

Article Title: Exploring potential additive effects of 5-fluorouracil, thymoquinone, and coenzyme Q10 triple therapy on colon cancer cells in relation to glycolysis and redox status modulation.
Article Snippet: Cell culture techniques and therapeutic intervention regimens All cell culture reagents were sourced from Thermo Fisher Scientific (MT, USA). .. Human HT29, SW480, and SW620 colon cancer cells (ATCC; VA, USA) were cultured in a humidified incubator (37°C and 5% CO2). ..

Article Title: Chemical composition, anticancer, antimicrobial activity of Aloysia citriodora Palau essential oils from four different locations in Palestine
Article Snippet: Human HS-5 stromal cultures were maintained in Iscove’s Modified Dulbecco’s Medium (IMDM; Wako, Japan), containing 10% FBS and 1% P/S. .. Human HT29 (ATCC; HTB-38), HCT 116 (ATCC CCL-247), SW620 [SW-620 (ATCC CCL-227), SW480 [SW-480] (ATCC CCL-228) cells were cultured in McCoy’s 5a Medium Modified, Wako, Japan) containing 10% FBS and 1% P/S. .. Human umbilical vein endothelial cells (HUVECs; ATCC, CRL-1730) were cultured at 37 °C/5% CO 2 on 0.1% gelatin (Wako Pure Chemicals)-coated culture plates (Falcon) in endothelial growth medium-2 (EGM-2; Lonza; cc4176).

other:

Article Title: Integrative proteomic profiling of tumor and plasma extracellular vesicles identifies a diagnostic biomarker panel for colorectal cancer
Article Snippet: Human: HT29 , ATCC , Cat# HTB-38.

Article Title: Protocol to identify covalent inhibitors targeting RhoA Cys16
Article Snippet: Human: HT29 (Wildtype/44Y/Female) , ATCC , #HTB-38.

Article Title: Tissue-dependent mechanosensing by cells derived from human tumors
Article Snippet: Human: HT29 , ATCC , HTB-38; RRID: CVCL_0320.

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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

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)).

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

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)).

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)).

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

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)).

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)).

Techniques: Activity Assay, Expressing, Control

PLSCR3 deficiency disrupts mitochondrial homeostasis in CRC cells. (A) Western blotting verified the reduction of PLSCR3 protein levels in HT29 cells. (B) RT‐qPCR analysis verified reduction of PLSCR3 mRNA levels normalized to GAPDH compared with negative control (NC) cells ( p < 0.0001, n = 3). (C) Representative JC‐1 staining (scale bar: 20 μ m) images showing red fluorescence (polarized mitochondria) and green fluorescence (depolarized mitochondria). (D) Quantification of the JC‐1 red/green fluorescence intensity ratio. Fluorescence intensity was quantified using ImageJ software as described in the section. (E) Real‐time oxygen consumption rate (OCR) profiles under sequential treatment with oligomycin, FCCP, and rotenone/Antimycin A. (F) Quantification of cytosolic mtDNA by qPCR using the D‐loop ratio in PLSCR3‐knockdown and control cells. Data are presented as mean ± SD from three independent experiments. Statistical significance was determined using an unpaired two‐tailed Student′s t ‐test for two‐group comparisons.

Journal: Human Mutation

Article Title: PLSCR3 Deficiency Triggers mtDNA‐Driven cGAS‐STING Activation to Potentiate Antitumor Immunity in Colorectal Cancer

doi: 10.1155/humu/8545428

Figure Lengend Snippet: PLSCR3 deficiency disrupts mitochondrial homeostasis in CRC cells. (A) Western blotting verified the reduction of PLSCR3 protein levels in HT29 cells. (B) RT‐qPCR analysis verified reduction of PLSCR3 mRNA levels normalized to GAPDH compared with negative control (NC) cells ( p < 0.0001, n = 3). (C) Representative JC‐1 staining (scale bar: 20 μ m) images showing red fluorescence (polarized mitochondria) and green fluorescence (depolarized mitochondria). (D) Quantification of the JC‐1 red/green fluorescence intensity ratio. Fluorescence intensity was quantified using ImageJ software as described in the section. (E) Real‐time oxygen consumption rate (OCR) profiles under sequential treatment with oligomycin, FCCP, and rotenone/Antimycin A. (F) Quantification of cytosolic mtDNA by qPCR using the D‐loop ratio in PLSCR3‐knockdown and control cells. Data are presented as mean ± SD from three independent experiments. Statistical significance was determined using an unpaired two‐tailed Student′s t ‐test for two‐group comparisons.

Article Snippet: The human colon carcinoma cell line HT29 (Cat. No. ATCC‐HTB‐38) and the mouse colon carcinoma cell line CT26.WT (Cat. No. CRL‐2638, ATCC, Virginia, United States) were maintained in RPMI 1640 medium (Cat. No. BC‐M‐017, Bio‐Channel, Jiangsu, China) supplemented with 10% fetal bovine serum.

Techniques: Western Blot, Quantitative RT-PCR, Negative Control, Staining, Fluorescence, Software, Knockdown, Control, Two Tailed Test

PLSCR3 deficiency activates mtDNA‐associated cGAS‐STING signaling. (A) 2 ′ ‐3 ′ ‐cGAMP quantification by ELISA in PLSCR3 knockdown (KD_PLSCR3) HT29 cells and normal controls (NCs) treated with Scramble or dideoxycytidine (DDC) ( p < 0.0001; ns = not significant; n = 3). (B, C) KD_PLSCR3 cells exhibited an elevation in IFN β and CXCL10 production measured by ELISA compared to NC ( p < 0.0001, n = 3). CRISPR‐mediated STING knockout (STING sg) completely abrogated these effects. (D) Western blot analysis showed upregulation of phosphorylated STING and innate immune effectors (IFIH1, ISG60, and ISG15 protein) in STING‐competent KD_PLSCR3 cells relative to GAPDH loading controls. (E) Normalized mRNA level quantification by qPCR demonstrated significant induction of interferon‐stimulated genes (ISGs) in STING‐competent KD_PLSCR3 cells compared with the other groups. Data in panels A–C and E are presented as mean ± SD. Statistical significance was determined using one‐way ANOVA followed by Tukey′s multiple‐comparisons post hoc test for multiple‐group comparisons.

Journal: Human Mutation

Article Title: PLSCR3 Deficiency Triggers mtDNA‐Driven cGAS‐STING Activation to Potentiate Antitumor Immunity in Colorectal Cancer

doi: 10.1155/humu/8545428

Figure Lengend Snippet: PLSCR3 deficiency activates mtDNA‐associated cGAS‐STING signaling. (A) 2 ′ ‐3 ′ ‐cGAMP quantification by ELISA in PLSCR3 knockdown (KD_PLSCR3) HT29 cells and normal controls (NCs) treated with Scramble or dideoxycytidine (DDC) ( p < 0.0001; ns = not significant; n = 3). (B, C) KD_PLSCR3 cells exhibited an elevation in IFN β and CXCL10 production measured by ELISA compared to NC ( p < 0.0001, n = 3). CRISPR‐mediated STING knockout (STING sg) completely abrogated these effects. (D) Western blot analysis showed upregulation of phosphorylated STING and innate immune effectors (IFIH1, ISG60, and ISG15 protein) in STING‐competent KD_PLSCR3 cells relative to GAPDH loading controls. (E) Normalized mRNA level quantification by qPCR demonstrated significant induction of interferon‐stimulated genes (ISGs) in STING‐competent KD_PLSCR3 cells compared with the other groups. Data in panels A–C and E are presented as mean ± SD. Statistical significance was determined using one‐way ANOVA followed by Tukey′s multiple‐comparisons post hoc test for multiple‐group comparisons.

Techniques: Enzyme-linked Immunosorbent Assay, Knockdown, CRISPR, Knock-Out, Western Blot

PLSCR3 deficiency enhances the sensitivity to immune cell–mediated killing in CRC. (A) Schematic of coculture system: PLSCR3 knockdown (KD_PLSCR3) HT29 cells or control were cocultured with human cord blood–derived NK cells at an E:T ratio of 5:1. (B, C) Phase‐contrast microscopy (scale bar: 50 μ m) showing morphological changes. Morphological analysis demonstrated decreased cell number in KD_PLSCR3 cells in the coculture condition ( p < 0.0001, n = 5). (D) Mouse graft carcinoma with control or PLSCR3 knockout (PLSCR3 sg) CT26 cells were treated with or without anti‐PD‐1 antibodies ( α PD‐1). Tumor growth curves (mm 3 ) were recorded at Days 5, 10, 15, and 20 after tumor cell inoculation in immunocompetent BALB/c mice. (E) At Day 20, terminal tumor weights in the mouse graft carcinoma were measured. (F–H) Flow cytometry analysis of CD4+ and CD8+ tumor‐infiltrating T cells. The representative cell populations of CD4+ and CD8+ are shown. (I, J) Flow cytometry and ELISA analysis of Granzyme B (GZMB) expression ( n = 5). Data are presented as mean ± SD. Tumor growth curves were analyzed using two‐way repeated‐measures ANOVA with post hoc multiple‐comparison tests. For multiple‐group comparisons, including panels I and J, one‐way ANOVA followed by Tukey′s post hoc test was used.

Journal: Human Mutation

Article Title: PLSCR3 Deficiency Triggers mtDNA‐Driven cGAS‐STING Activation to Potentiate Antitumor Immunity in Colorectal Cancer

doi: 10.1155/humu/8545428

Figure Lengend Snippet: PLSCR3 deficiency enhances the sensitivity to immune cell–mediated killing in CRC. (A) Schematic of coculture system: PLSCR3 knockdown (KD_PLSCR3) HT29 cells or control were cocultured with human cord blood–derived NK cells at an E:T ratio of 5:1. (B, C) Phase‐contrast microscopy (scale bar: 50 μ m) showing morphological changes. Morphological analysis demonstrated decreased cell number in KD_PLSCR3 cells in the coculture condition ( p < 0.0001, n = 5). (D) Mouse graft carcinoma with control or PLSCR3 knockout (PLSCR3 sg) CT26 cells were treated with or without anti‐PD‐1 antibodies ( α PD‐1). Tumor growth curves (mm 3 ) were recorded at Days 5, 10, 15, and 20 after tumor cell inoculation in immunocompetent BALB/c mice. (E) At Day 20, terminal tumor weights in the mouse graft carcinoma were measured. (F–H) Flow cytometry analysis of CD4+ and CD8+ tumor‐infiltrating T cells. The representative cell populations of CD4+ and CD8+ are shown. (I, J) Flow cytometry and ELISA analysis of Granzyme B (GZMB) expression ( n = 5). Data are presented as mean ± SD. Tumor growth curves were analyzed using two‐way repeated‐measures ANOVA with post hoc multiple‐comparison tests. For multiple‐group comparisons, including panels I and J, one‐way ANOVA followed by Tukey′s post hoc test was used.

Techniques: Knockdown, Control, Derivative Assay, Microscopy, Knock-Out, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Expressing, Comparison

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adenocarcinoma derived cell line from human ht29 (ATCC)

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1) Product Images from "Condensin upregulation induces intestinal stem cell senescence, driving cell death and increased intestinal permeability"

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