Journal: Journal of Translational Medicine

Article Title: Development of patient-derived xenograft models from a spontaneously immortal low-grade meningioma cell line, KCI-MENG1

doi: 10.1186/s12967-015-0596-8

Figure Lengend Snippet: Immunostaining of original tumor, low passage, and high passage KCI-MENG1 cells. The original patient-derived tumor ( top row ) showed moderate and patchy immunoreactivity for epithelial membrane antigen (EMA); strong and diffuse immunostaining for progesterone receptor (PR); and a Ki-67 proliferative index of 2–3%. There was also strong immunostaining for N-cadherin and vimentin. KCI-MENG1-LP cells ( middle row ) and KCI-MENG1-HP cells ( bottom row ) maintained expression of EMA, N-cadherin, and vimentin but had significantly reduced PR expression compared to the original tumor. Whereas Ki-67 labeling was found in only a small number of cells in the original tumor and low passage cells, it was positive in virtually all P84 cells. Scale bar 50 µm.

Article Snippet: Primary antibodies used targeted the following proteins: EMA (cat.#247M-94), PR (cat.#323R-14), Ki-67 (cat.#275R-14), vimentin (cat.#347R-14; all from CellMarque, Rocklin, CA, USA), and N-cadherin (cat.#NBP1-48309, Novus Biologicals, Littleton, CO, USA).

Techniques: Immunostaining, Derivative Assay, Membrane, Expressing, Labeling

Journal: Journal of Translational Medicine

Article Title: Development of patient-derived xenograft models from a spontaneously immortal low-grade meningioma cell line, KCI-MENG1

doi: 10.1186/s12967-015-0596-8

Figure Lengend Snippet: Immunostaining of original patient tumor, low and high passage KCI-MENG1 cells, and subcutaneous xenograft tumor. The original patient-derived tumor showed moderate immunoreactivity for E-cadherin which was maintained in all in vitro and in vivo models. Scale bar 50 µm.

Article Snippet: Primary antibodies used targeted the following proteins: EMA (cat.#247M-94), PR (cat.#323R-14), Ki-67 (cat.#275R-14), vimentin (cat.#347R-14; all from CellMarque, Rocklin, CA, USA), and N-cadherin (cat.#NBP1-48309, Novus Biologicals, Littleton, CO, USA).

Techniques: Immunostaining, Derivative Assay, In Vitro, In Vivo

Journal: Journal of Translational Medicine

Article Title: Development of patient-derived xenograft models from a spontaneously immortal low-grade meningioma cell line, KCI-MENG1

doi: 10.1186/s12967-015-0596-8

Figure Lengend Snippet: Meningioma cell lines reported in the literature

Article Snippet: Primary antibodies used targeted the following proteins: EMA (cat.#247M-94), PR (cat.#323R-14), Ki-67 (cat.#275R-14), vimentin (cat.#347R-14; all from CellMarque, Rocklin, CA, USA), and N-cadherin (cat.#NBP1-48309, Novus Biologicals, Littleton, CO, USA).

Techniques: Southern Blot, Injection, Expressing, TRAP Assay, Activity Assay, Real-time Polymerase Chain Reaction

Journal: Journal of Translational Medicine

Article Title: Development of patient-derived xenograft models from a spontaneously immortal low-grade meningioma cell line, KCI-MENG1

doi: 10.1186/s12967-015-0596-8

Figure Lengend Snippet: Human meningioma mouse xenograft model KCI-MENG1-LPSX generated with the spontaneously immortal cell line KCI-MENG1-LP. Tumors from immunocompromised SCID mice were dissected ( a ) and the derivative cell line KCI-MENG1-LPSX CL was generated. The H&E staining of the mouse tumor revealed a pattern of moderately cellular meningothelial cells similar to the original patient tumor ( b ). The KCI-MENG1-LPSX CL cells were composed of the round-shaped cells similar to the high passage parent cell line KCI-MENG1-HP ( c ). The EMA, PR, and N-cadherin IHC of the mouse tumor highly resembled the original patient-derived tumor ( d top row ). The vimentin- and Ki-67-stained cells in the mouse tumor tissue were markedly more abundant and more intensely stained than in the original tumor ( d top row ). KCI-MENG1-LPSX CL cells displayed the same patterns of immunostaining as the high passage parent cell line KCI-MENG1-HP, including the loss of PR staining ( d bottom row ). Scale bar 50 µm.

Article Snippet: Primary antibodies used targeted the following proteins: EMA (cat.#247M-94), PR (cat.#323R-14), Ki-67 (cat.#275R-14), vimentin (cat.#347R-14; all from CellMarque, Rocklin, CA, USA), and N-cadherin (cat.#NBP1-48309, Novus Biologicals, Littleton, CO, USA).

Techniques: Generated, Staining, Derivative Assay, Immunostaining

Journal: Journal of Molecular Endocrinology

Article Title: Role of reduced expression of SMAD4 in papillary thyroid carcinoma

doi: 10.1677/jme-10-0044

Figure Lengend Snippet: Figure 7 Migratory behavior of TPC-1 and BCPAP. (A) Wound-healing assay was performed in confluent monolayers for 24 h, and the extension of the area colonized by the cells was estimated using Axio Vision, Zeiss software (** indicates a statistical significance (Student’s t-test, P!0.001) compared to FRTL-5). (B) Wound-healing assay in TPC-1 and BCPAP overexpressing SMAD4 wt, untreated or treated with 10 ng/ml of TGFb1 for 24 h (** indicates a high statistical significance (Student’s t-test, P!0.001) compared to empty vector control cells). Images are representative of three separate experiments. (C) Whole protein lysates (40 mg/lane) from TPC-1, BCPAP, FTC133, and FRTL-5 control cells, MCF7, and HEK293, were analyzed by western blot using antibodies against E-cadherin and N-cadherin. (D) Western blot of whole protein lysates (40 mg/lane) from TPC-1 and BCPAP transiently transfected with pEGFPC3-SMAD4 wt or pEGFPC3 empty vector performed using an antibody against E-cadherin. Bottom, densitometric evaluation of the E-cadherin bands normalized to the levels of b-actin (** indicates a high statistical significance (Student’s t-test), P!0.001).

Article Snippet: Phenylmethylsulfonyl fluoride (PMSF), protease inhibitor cocktail containing 1 mg/ml leupeptin, 2.5 mg/ml aprotinin, and 1 mg/ml benzamide hydrochloride, and phosphatase inhibitor cocktail were purchased from Sigma–Aldrich, Co. Antibodies to TbRI and TbRII, green fluorescent protein (GFP), SMURF1 (H60), SMAD7 (H79), SMAD4 (B8), SMAD4 (H552), N-cadherin (H63), b-actin (C4), and Lamin B (C20) were obtained from Santa Cruz Biotechnology (Santa Cruz, Journal of Molecular Endocrinology (2010) 45, 229–244 CA, USA).

Techniques: Wound Healing Assay, Software, Plasmid Preparation, Control, Western Blot, Transfection

Journal: International Journal of Molecular Sciences

Article Title: Depletion of Lipocalin 2 (LCN2) in Mice Leads to Dysbiosis and Persistent Colonization with Segmented Filamentous Bacteria

doi: 10.3390/ijms222313156

Figure Lengend Snippet: Comparative analysis of the intestine macroscopical parameters and expression of Lcn2 in wild type (n = 4) and Lcn2 -/- mice (n = 4 or 5). ( A ) Evaluation of the body weight of male animals at the same age (left). Corresponding ceca were prepared and weighed (middle). Ratio of body to cecal weight (right). ( B ) Gross macroscopical and graphical comparison of the total small and large intestinal lengths measured (n = 4 or 5). ( C ) Quantitative analysis of Lcn2 mRNA (upper) and LCN2 protein expression (lower) in the distal small intestine (left) cecum (middle), and distal colon (right) of wild type ( Lcn2 +/+ ) and Lcn2 null mice (each n = 4 or 5). Relative mRNA expression of respective genes was done by RT-qPCR and normalized to Gapdh or β-actin . Equal protein loading was demonstrated by re-probing the membranes with a specific antibody directed against GAPDH or β-actin. E-cadherin was probed as a marker for epithelial-to-mesenchymal transition (EMT). Primers used are given in . Statistical evaluation in ( A – C ) was performed by an unpaired t -test, whereby the difference of two samples from a level of p < 0.05 is considered statistically significant (*: p < 0.05, **: p < 0.01, ***: p < 0.001).

Article Snippet: For detection of individual proteins, the primary antibodies for LCN2/NGAL (AF3508, R & D Systems, Abingdon, UK), GAPDH (6C5) (sc-32233, Santa Cruz, Biotech, Santa Cruz, CA, USA), β-actin (A5441, Sigma–Aldrich, MI, USA), and E-cadherin (AP20195PU-N, OriGene, Rockville, MD, USA) were diluted in 2.5% ( w / v ) non-fat milk powder in TBST.

Techniques: Expressing, Comparison, Quantitative RT-PCR, Marker