Journal: Molecular Oncology

Article Title: SNAIL1 combines competitive displacement of ASCL2 and epigenetic mechanisms to rapidly silence the EPHB3 tumor suppressor in colorectal cancer

doi: 10.1016/j.molonc.2014.08.016

Figure Lengend Snippet: Snail1‐HA completely disables the EPHB3 enhancer. (A–C) ChIP analyses of Snail1‐HA (A), TCF7L2 (B) and H3K4me1 (C) occupancy at the EPHB3 locus in derivatives of LS174T cells 96 h after Snail1‐HA induction. n ≥ 3. ...

Article Snippet: Cell culture The CRC cell lines LS174T (CLS #300392) and HT29 (CLS #300215) were obtained from the Cell Line Service culture collection (DKFZ, Heidelberg) and ATCC, respectively.

Techniques:

Journal: Molecular Oncology

Article Title: SNAIL1 combines competitive displacement of ASCL2 and epigenetic mechanisms to rapidly silence the EPHB3 tumor suppressor in colorectal cancer

doi: 10.1016/j.molonc.2014.08.016

Figure Lengend Snippet: Snail1‐HA engages HDACs and LSD1 to repress EPHB3. (A,B) ChIP analyses of H3ac, H3K4me3 (A) and HDAC1 occupancy (B) in derivatives of LS174T cells. n ≥ 3. (C) qRT‐PCR analyzing HDAC1, HDAC2 and EPHB3 expression ...

Article Snippet: Cell culture The CRC cell lines LS174T (CLS #300392) and HT29 (CLS #300215) were obtained from the Cell Line Service culture collection (DKFZ, Heidelberg) and ATCC, respectively.

Techniques: Quantitative RT-PCR, Expressing

Journal: Molecular Oncology

Article Title: SNAIL1 combines competitive displacement of ASCL2 and epigenetic mechanisms to rapidly silence the EPHB3 tumor suppressor in colorectal cancer

doi: 10.1016/j.molonc.2014.08.016

Figure Lengend Snippet: Snail1‐HA expression interferes with marker gene expression of intestinal epithelial stem cells and self‐renewal capacity. (A) qRT‐PCR analyses of the genes shown in derivatives of LS174T cells. Expression (expr.) levels are shown ...

Article Snippet: Cell culture The CRC cell lines LS174T (CLS #300392) and HT29 (CLS #300215) were obtained from the Cell Line Service culture collection (DKFZ, Heidelberg) and ATCC, respectively.

Techniques: Expressing, Marker, Gene Expression, Quantitative RT-PCR

Journal: Molecular Oncology

Article Title: SNAIL1 combines competitive displacement of ASCL2 and epigenetic mechanisms to rapidly silence the EPHB3 tumor suppressor in colorectal cancer

doi: 10.1016/j.molonc.2014.08.016

Figure Lengend Snippet: Sustained expression of EPHB3 interferes with Snail1‐HA‐induced EMT. (A) Scheme illustrating the construction of LS174T CRC cell derivatives. In a first step, LS174T cells were tranduced with a retroviral vector for Dox‐inducible ...

Article Snippet: Cell culture The CRC cell lines LS174T (CLS #300392) and HT29 (CLS #300215) were obtained from the Cell Line Service culture collection (DKFZ, Heidelberg) and ATCC, respectively.

Techniques: Expressing, Retroviral, Plasmid Preparation

Journal: Molecular Oncology

Article Title: SNAIL1 combines competitive displacement of ASCL2 and epigenetic mechanisms to rapidly silence the EPHB3 tumor suppressor in colorectal cancer

doi: 10.1016/j.molonc.2014.08.016

Figure Lengend Snippet: EPHB3 counteracts Snail1‐HA‐induced EMT in vivo. Representative images of serial sections of xenograft tumors formed by LS174T control cells or derivatives expressing Snail1‐HA and Snail1‐HA in the continuous presence ...

Article Snippet: Cell culture The CRC cell lines LS174T (CLS #300392) and HT29 (CLS #300215) were obtained from the Cell Line Service culture collection (DKFZ, Heidelberg) and ATCC, respectively.

Techniques: In Vivo, Control, Expressing

Journal: mAbs

Article Title: Generation and in vivo characterization of a novel high-affinity human antibody targeting carcinoembryonic antigen

doi: 10.1080/19420862.2023.2217964

Figure Lengend Snippet: In vitro characterization of new anti-CEA antibodies. (a) SPR sensorgram of G9, F7, and F4 in a scFv format. K D values were calculated to be 640 nM, 50 nM, and 7.7 nM, respectively. (b) Flow cytometry analysis with the new antibodies in IgG format on CEA-expressing CT26 cells and on CEA-negative CT26 wild-type cells. (c) Immunofluorescence staining with IgG formats on the human colon adenocarcinoma xenograft LS174T. Anti-CEA antibodies were detected in green. Blood vessels were detected by CD31 staining (red). 20× magnification, scale bars = 100 μm.

Article Snippet: Immunofluorescence analysis was performed on LS174T and HT-29 xenografts harvested from mice, on patient-derived colon cancer samples, and on a human tissue microarray (Amsbio, T6235700–5).

Techniques: In Vitro, Flow Cytometry, Expressing, Immunofluorescence, Staining

Journal: mAbs

Article Title: Generation and in vivo characterization of a novel high-affinity human antibody targeting carcinoembryonic antigen

doi: 10.1080/19420862.2023.2217964

Figure Lengend Snippet: Ex vivo immunofluorescence-based biodistribution analysis. Immunofluorescence analysis assessed tumor targeting of new anti-CEA antibodies in IgG format. Two hundred micrograms of IgG-FITC were injected intravenously into LS174T-bearing mice. Tumors were excised 24 hours after injection. IgG-FITC was detected in green; blood vessels were detected through CD31 staining (red). 20× magnification, scale bars = 100 μm.

Article Snippet: Immunofluorescence analysis was performed on LS174T and HT-29 xenografts harvested from mice, on patient-derived colon cancer samples, and on a human tissue microarray (Amsbio, T6235700–5).

Techniques: Ex Vivo, Immunofluorescence, Injection, Staining

Journal: mAbs

Article Title: Generation and in vivo characterization of a novel high-affinity human antibody targeting carcinoembryonic antigen

doi: 10.1080/19420862.2023.2217964

Figure Lengend Snippet: Quantitative biodistribution with radiolabeled anti-CEA antibodies in diabody format. Quantitative biodistribution analysis of radio iodinated anti-CEA diabodies in BALB/c nude mice bearing subcutaneous LS174T colon adenocarcinomas. Organs were harvested 24 hours after intravenous injection, and radioactivity was quantified. Results are shown as the percentage of injected dose per gram (ID/g (%)). Error bars = SEM; n = 4.

Article Snippet: Immunofluorescence analysis was performed on LS174T and HT-29 xenografts harvested from mice, on patient-derived colon cancer samples, and on a human tissue microarray (Amsbio, T6235700–5).

Techniques: Injection, Radioactivity

Journal: Glycobiology

Article Title: Neutralization of pH in the Golgi apparatus causes redistribution of glycosyltransferases and changes in the O-glycosylation of mucins.

doi: 10.1093/glycob/11.8.633

Figure Lengend Snippet: Fig. 7. Subcellular fractionation of HeLa (A–C) and LS 174T (D–F) cells for assay of glycosyltransferase relocalization on NH4Cl treatment. Cells were homogenized in buffered KCl by passage through syringe needles, debris was pelleted, and the obtained postnuclear supernatant was fractionated by ultracentrifugation on a linear Nycodenz gradient. The nine fractions, recovered from the top to the bottom, were subjected to tabletop ultracentrifugation, and the obtained pellets were reduced and analyzed by western blots of SDS–PAGE, assaying calnexin (A and D), GalNAc-T2/VSV-G (B), endogenous GalNAc-T2 (E) and Gal-T1 (C and F), and developed by the ECL method. In addition to images of the western blots, where the upper in every pair is from control cells and the lower from NH4Cl-treated cells, video densitometry measurements are shown. Open circles, control cells; filled-in circles, NH4Cl-treated cells. The difference in calnexin staining between control and treated cells in A, fraction 8, is not significant.

Article Snippet: The MUC2-producing colon adenocarcinoma cell line LS 174T (ATCC CL 188) (Asker et al., 1995), and monolayer HeLa cells (ATCC CCL 185), stable expressing VSV-Gtagged GalNAc-T2 (Röttger et al., 1998), myc-tagged NAGT I (Nilsson et al., 1993), and GFP-tagged KDEL receptor, were cultivated as described.

Techniques: Fractionation, Western Blot, SDS Page, Control, Staining

Journal: Glycobiology

Article Title: Neutralization of pH in the Golgi apparatus causes redistribution of glycosyltransferases and changes in the O-glycosylation of mucins.

doi: 10.1093/glycob/11.8.633

Figure Lengend Snippet: Fig. 8. Effect of NH4Cl treatment on the Golgi structure in LS 174T cells. Immuno-EM of ultrathin cryosections of control cells (A) and cells treated with 25 mM NH4Cl for 40 h (B). Bar, 200 nm.

Article Snippet: The MUC2-producing colon adenocarcinoma cell line LS 174T (ATCC CL 188) (Asker et al., 1995), and monolayer HeLa cells (ATCC CCL 185), stable expressing VSV-Gtagged GalNAc-T2 (Röttger et al., 1998), myc-tagged NAGT I (Nilsson et al., 1993), and GFP-tagged KDEL receptor, were cultivated as described.

Techniques: Control

Journal: Glycobiology

Article Title: Neutralization of pH in the Golgi apparatus causes redistribution of glycosyltransferases and changes in the O-glycosylation of mucins.

doi: 10.1093/glycob/11.8.633

Figure Lengend Snippet: Fig. 9. Gas chromatograms of released mucin oligosaccharides. O-linked oligosaccharides were released from purified insoluble mucins (MUC2) of LS 174T cells by β-elimination and fractionated into neutral, sialylated, and sulfated fractions. Gas chromatograms of permethylated neutral (A and C) and sialylated (B and D) oligosaccharides released from mucins of control (A and B) and NH4Cl-treated (C and D) cells. Designations on the peaks refer to Figure 10.

Article Snippet: The MUC2-producing colon adenocarcinoma cell line LS 174T (ATCC CL 188) (Asker et al., 1995), and monolayer HeLa cells (ATCC CCL 185), stable expressing VSV-Gtagged GalNAc-T2 (Röttger et al., 1998), myc-tagged NAGT I (Nilsson et al., 1993), and GFP-tagged KDEL receptor, were cultivated as described.

Techniques: Purification, Control

Journal: Hormones & Cancer

Article Title: Targeting the Mitotic Checkpoint to Kill Tumor Cells

doi: 10.1007/s12672-010-0059-x

Figure Lengend Snippet: Conditional inactivation of Mps1 causes inactivation of the mitotic checkpoint and tumor cell death. a Left LS174T-TetRMps1, treated without (−) and with (+) doxycycline (dox) for 3 days, were immunoblotted for Mps1 and α-tubulin. Right colony formations of LS174T-TetRMps1 cells treated with and without dox for 11 days. b FACS analysis of LS174T-TetRMps1 cells treated with or without dox for 3 days and 1 μM taxol for 18 h. The percentage of mitotic cells was measured as the fraction of cells with 4N DNA content that were positive for MPM2

Article Snippet: LS174T cells were grown in DMEM (Lonza) with 10% Tet-approved FCS (Clontech), supplemented with pen/strep (Invitrogen) and ultraglutamine (Lonza).

Techniques:

Journal: Hormones & Cancer

Article Title: Targeting the Mitotic Checkpoint to Kill Tumor Cells

doi: 10.1007/s12672-010-0059-x

Figure Lengend Snippet: Conditional inactivation of Mps1 causes severe defects in chromosome segregation and severe aneuploidy. a LS174T-TetRMps1 cells were treated with or without dox for 3 days and fixed after 90 min of MG132 treatment. No alignment defects , mild alignment defects , or severe alignment defects indicate mitotic cells with zero, one to five, or more than five chromosomes not aligned on the metaphase plate, respectively. b Chromosome spreads were performed on cells after 4 days of treatment with or without dox. At least 60 chromosome spreads were counted per condition

Article Snippet: LS174T cells were grown in DMEM (Lonza) with 10% Tet-approved FCS (Clontech), supplemented with pen/strep (Invitrogen) and ultraglutamine (Lonza).

Techniques: