Journal: Journal of Virology

Article Title: Molecular Determinants of Species Specificity in the Coronavirus Receptor Aminopeptidase N (CD13): Influence of N-Linked Glycosylation

doi: 10.1128/jvi.75.20.9741-9752.2001

Figure Lengend Snippet: FIG. 2. Mutations in amino acids 288 to 295 of hAPN blocked HCoV-229E infection. CMT93 cell lines were generated by transfec- tion with wt-hAPN (A and B), p6-hAPN (C and D), or pCi-neo (E and F), and the hAPN-expressing lines (A to D) were sorted by FACS for similar expression levels of hAPN protein. hAPN protein was shown by indirect IFA with MAb WM47 (A, C, and E) and infection by HCoV- 229E was shown by indirect IFA, 22 h postinoculation, with anti-spike glycoprotein MAb 511H.6 (B, D, and F).

Article Snippet: The BHK-21 line of baby hamster kidney fibroblasts (from Robert Garcea, University of Colorado Health Sciences Center, Denver), CMT93 cells (ATCC), swine testicle cells (from David Brian, University of Tennessee, Knoxville), and Felis catus whole fetus (FCWF) cells (from Niels Pedersen, University of California at Davis) were grown in Dulbecco’s modified MEM (DMEM) supplemented with 10% heat-inactivated FBS (GIBCO BRL).

Techniques: Infection, Generated, Expressing

Journal: Journal of Virology

Article Title: Molecular Determinants of Species Specificity in the Coronavirus Receptor Aminopeptidase N (CD13): Influence of N-Linked Glycosylation

doi: 10.1128/jvi.75.20.9741-9752.2001

Figure Lengend Snippet: FIG. 4. Addition of an N-linked glycosylation site at amino acid 291 of hAPN blocked HCoV-229E receptor activity. Cell lysates isolated from stably transfected CMT93 cell lines that were sorted by FACS to express similar levels of hAPN were analyzed by immunoblotting. Cell lysates were isolated from cells expressing wt-hAPN (lane 1), p6-hAPN (lane 2), hAPN/N291KT (lane 3), hAPN/E291KT (lane 4), hAPN/N291KQ (lane 5), or a line created with the empty expression vector pCi-neo (lane 6). The cells were inoculated with HCoV-229E at an MOI of 2, and the cell lysates were prepared 21.5 h postinoculation. The proteins were separated by SDS–10% PAGE and transferred to Immobilon-P, and HCoV-229E nucleocapsid (N) protein expression (arrow) was shown using polyclonal goat antiserum to HCoV-229E.

Article Snippet: The BHK-21 line of baby hamster kidney fibroblasts (from Robert Garcea, University of Colorado Health Sciences Center, Denver), CMT93 cells (ATCC), swine testicle cells (from David Brian, University of Tennessee, Knoxville), and Felis catus whole fetus (FCWF) cells (from Niels Pedersen, University of California at Davis) were grown in Dulbecco’s modified MEM (DMEM) supplemented with 10% heat-inactivated FBS (GIBCO BRL).

Techniques: Glycoproteomics, Activity Assay, Isolation, Stable Transfection, Transfection, Western Blot, Expressing, Plasmid Preparation

Journal: Journal of Virology

Article Title: Molecular Determinants of Species Specificity in the Coronavirus Receptor Aminopeptidase N (CD13): Influence of N-Linked Glycosylation

doi: 10.1128/jvi.75.20.9741-9752.2001

Figure Lengend Snippet: FIG. 6. Binding of HCoV-229E was inhibited by the addition of a glycosylation signal at amino acid 291 of hAPN. CMT93 cell lines that had similar levels of hAPN surface expression were mixed with 3H-labeled virus, the cells were washed and lysed, and 3H was counted as described in Materials and Methods. The average 3H-labeled virus bound by the wt-hAPN was 1.38 104 cpm and was defined as 100% binding. The average total counts bound by cells transfected with pCi-neo was defined as 0% binding. The average of the other cell lines was reported as a percentage of wild-type binding activity, and the error bars represent the standard deviation of the three replicates.

Article Snippet: The BHK-21 line of baby hamster kidney fibroblasts (from Robert Garcea, University of Colorado Health Sciences Center, Denver), CMT93 cells (ATCC), swine testicle cells (from David Brian, University of Tennessee, Knoxville), and Felis catus whole fetus (FCWF) cells (from Niels Pedersen, University of California at Davis) were grown in Dulbecco’s modified MEM (DMEM) supplemented with 10% heat-inactivated FBS (GIBCO BRL).

Techniques: Binding Assay, Glycoproteomics, Expressing, Labeling, Virus, Transfection, Activity Assay, Standard Deviation

Journal: Journal of Virology

Article Title: Molecular Determinants of Species Specificity in the Coronavirus Receptor Aminopeptidase N (CD13): Influence of N-Linked Glycosylation

doi: 10.1128/jvi.75.20.9741-9752.2001

Figure Lengend Snippet: FIG. 7. The influence of a glycosylation signal at N818 of hAPN and p6-hAPN on receptor activity for serogroup 1 coronaviruses of humans, pigs, and cats. Nucleocapsid (N) proteins (arrows) were identified by immunoblotting with goat polyclonal antisera to HCoV-229E (A) or feline polyclonal antisera to FIPV that cross-reacts with TGEV (B and C). FCWF (lane 1) or CMT93 cell lines that express wt-hAPN (lane 2), p6-hAPN (lane 3), hAPN/N818E (lane 4), p6-hAPN/N818E (lane 5), and p6-hAPN/T820E (lane 6) or cell lines created with the empty expression vector pCi-neo (lane 7) were inoculated with HCoV-229E (A), TGEV (B), or FECoV 79-1863 (C).

Article Snippet: The BHK-21 line of baby hamster kidney fibroblasts (from Robert Garcea, University of Colorado Health Sciences Center, Denver), CMT93 cells (ATCC), swine testicle cells (from David Brian, University of Tennessee, Knoxville), and Felis catus whole fetus (FCWF) cells (from Niels Pedersen, University of California at Davis) were grown in Dulbecco’s modified MEM (DMEM) supplemented with 10% heat-inactivated FBS (GIBCO BRL).

Techniques: Glycoproteomics, Activity Assay, Western Blot, Expressing, Plasmid Preparation

Journal:

Article Title: An ?- E-catenin gene trap mutation defines its function in preimplantation development

doi:

Figure Lengend Snippet: β-Galactosidase expression in α-E-cateninGT1 heterozygous embryos. All plates show specimens stained for detection of β-galactosidase. (A) Whole mount E12 embryo. (B) Mesonephros and gonad stained after dissection from an E12 embryo. (C) A lung primordium from the same embryo. (D) Detail of the head region of a sagittal section of an E13 embryo after β-galactosidase staining. (E) Whole mount expanded blastocyst. e, Inner ear; eb, epithelial branches in the lung; g, gonad; icm, inner cell mass; m, mesenchyme of the lung; md, mesonephric duct; ms, mesencephalon; mt, mesonephric tubules; pmd, paramesonephric duct; so, somites; t, telencephalon; tr, trophoblast. Arrowheads in E point to the junctions between trophoblast cells. Open arrowhead in D points to the ventricular layer of the prospective telencephalic cortical region, and filled arrowhead points to the nasal epithelium.

Article Snippet: CMT cells used as controls are a mouse rectum carcinoma cell line with epithelial morphology (ATCC CCL 223).

Techniques: Expressing, Staining, Dissection

Journal: Oncology Letters

Article Title: Fatty acids inhibit anticancer effects of 5-fluorouracil in mouse cancer cell lines

doi: 10.3892/ol.2017.6190

Figure Lengend Snippet: Effect of LA or EA on 5-FU-induced reduction of cancer cell viability. CT26 cells were treated with (A) LA (50 µg/ml) or (B) EA (20 µg/ml), administered concurrently with 5-FU (1 µg/ml). LL2 cells were treated with (C) LA (50 µg/ml) or (D) EA (20 µg/ml), administered concurrently with 5-FU (1 µg/ml). CMT93 cells were treated with (E) LA (50 µg/ml) or (F) EA (20 µg/ml), administered concurrently with 5-FU (1 µg/ml). (G) LA (50 µg/ml) (H) or EA (20 µg/ml) was administered 24 h prior to 5-FU treatment (1 µg/ml) in 3 cancer cell lines. Data are expressed as the mean ± standard deviation. LA, linoleic acid; EA, elaidic acid; 5-FU, 5-fluorouracil; Mock, vehicle treated with 70% ethanol.

Article Snippet: The mouse rectal carcinoma CMT93 cell line and the mouse lung cancer LL2 cell line were purchased from DS Pharma Biomedical Co., Ltd. (Osaka, Japan).

Techniques: Standard Deviation

Journal: PLoS Neglected Tropical Diseases

Article Title: 2,4-Diaminothieno[3,2- d ]pyrimidines, a new class of anthelmintic with activity against adult and egg stages of whipworm

doi: 10.1371/journal.pntd.0006487

Figure Lengend Snippet: Summary of the cytotoxicity in a mouse epithelial cell line of the DATP series. Mouse CMT-93 rectal epithelial cells were used for this assay. Maximum tested concentration was 100 μM. n = 8, error range (in parentheses) shows 95% confidence interval. EC 50 values in the adult Trichuris paralysis assay are shown for comparison.

Article Snippet: The mouse rectal epithelial cell line CMT-93 (LGC Promochem, Teddington, United Kingdom) was used for these studies.

Techniques: Concentration Assay, Comparison