Journal: Molecular Biology of the Cell

Article Title: Synergistic interaction between the fibroblast growth factor and bone morphogenetic protein signaling pathways in lens cells

doi: 10.1091/mbc.E15-02-0117

Figure Lengend Snippet: Expression of mRNA for the indicated test gene as measured by qRT-PCR (TaqMan), using GAPDH for normalization.

Article Snippet: Thermal cycling conditions were as follows: 95 °C for 10 min, 50 cycles of 95 °C/10 s and 60 °C/20 s, and 40 °C for 30 s. PCRs contained twofold dilutions of cDNA template, TaqMan Universal PCR Master Mix, and TaqMan Assay Mix (for Id1 , FGFR1 , FGFR3 , BMP7 , BMP4 , and GAPDH , assay IDs were, respectively, Gg03337774_g1, Gg03340351_m1, Gg03340329_m1, Gg03310499_m1, Gg03348675_m1, and Gg03346982_m1).

Techniques: Expressing

Journal: Molecular Biology of the Cell

Article Title: Synergistic interaction between the fibroblast growth factor and bone morphogenetic protein signaling pathways in lens cells

doi: 10.1091/mbc.E15-02-0117

Figure Lengend Snippet: FGF specifically increases expression of the BMP signaling reporter BRE-Luc in lens cells. (A–E) DCDMLs were transfected with plasmids encoding BRE-Luc (A–C), SBE4-Luc (D), or 5′ Del 6 Id1-Luc (E) and cultured with no additions (control), 5 ng/ml BMP4, 10 ng/ml FGF2, 1 ng/ml FGF2, 4 ng/ml TGFβ1, or 50 nM TPA for 22 h (A, B, D, E), or 2 h (C). Where indicated, the FGFR inhibitor PD173074 (PD) was also present. Cultures were either fixed and immunostained with anti-luciferase antibodies (A, D, E) or lysed and equal amount of total cell protein analyzed by Western blot using the same antibody (B, C). Representative of three or more independent experiments. (F) Experiments in which a plasmid encoding constitutively expressed EGFP was cotransfected with 5′ Del 6 Id1-Luc at a ratio of 1:4 demonstrated equal levels of expression of EGFP when normalized to either total protein or tubulin, indicating that the lack of expression of 5′ Del 6 Id1-Luc in E was not due to a lack of cell transfection.

Article Snippet: Thermal cycling conditions were as follows: 95 °C for 10 min, 50 cycles of 95 °C/10 s and 60 °C/20 s, and 40 °C for 30 s. PCRs contained twofold dilutions of cDNA template, TaqMan Universal PCR Master Mix, and TaqMan Assay Mix (for Id1 , FGFR1 , FGFR3 , BMP7 , BMP4 , and GAPDH , assay IDs were, respectively, Gg03337774_g1, Gg03340351_m1, Gg03340329_m1, Gg03310499_m1, Gg03348675_m1, and Gg03346982_m1).

Techniques: Expressing, Transfection, Cell Culture, Control, Luciferase, Western Blot, Plasmid Preparation

Journal: Oncology Letters

Article Title: Expression of TRPM8 in human reactive lymphoid tissues and mature B-cell neoplasms

doi: 10.3892/ol.2018.9386

Figure Lengend Snippet: Immunohistochemistry of TRPM8 and lymphocyte markers in tonsils and mature B-cell neoplasms. (A) The CD23 immunostain recognizes four different zones of a tonsillar lymphoid follicle, namely, the MaZ, outer zone (*), LZ, and DZ. MZ: SYM / MZ ; IF: Interfollicular area (×40). (B) The TRPM8 immunostain in the same lymphoid follicle as (A). There are many TRPM8 + cells in the light zone and SYM/MZ (×40). (C) TRPM8 + cells in the light zone of a tonsillar lymphoid follicle (×200). (D) TRPM8 + cells in SYM/MZ (×400). (E) Double immunostain of IRF4/MUM-1 and TRPM8 in the tonsillar germinal center using digital photomicrographs. The nuclei of IRF4/MUM-1 + cells are shown as blue in a digital photo. The same tissue section was immunostained with TRPM8 and IRF4/MUM-1 + TRPM8 + cells, shown as red in the photo. The first photo was overlaid with the second photo. IRF4/MUM-1 + TRPM8 + cells are recognized as cells having a black-colored nucleus and red-colored cytoplasm (×200). (F) TRPM8 + neoplastic cells in plasma cell myeloma; immunostaining score=4 (×200). (G) TRPM8 + cells in lymphoplasmacytic lymphoma; immunostaining score=2 (×200). (H) TRPM8 + cells in mantle cell lymphoma; immunostaining score=0 (×200). All sections were counterstained with hematoxylin. SYM, lymphoepithelial symbiosis; MZ, marginal zone; TRPM8, transient receptor potential melastatin 8; DZ, dark zonel LZ, light zone; MaZ, mantle zone; IF: Interfollicular area.

Article Snippet: Immunohistochemistry was performed using a rabbit polyclonal anti-human TRPM8 antibody [Alomone, Jerusalem, Israel; diluted titer of 1/200; recognizes the peptide SDVD GTTYDFAHC corresponding to amino acid residues 917–929 in the 3rd extracellular loop of human TRPM8 (Accession Q7Z2W7)] ( ) as a primary antibody.

Techniques: Immunohistochemistry, Immunostaining

Journal: Oncology Letters

Article Title: Expression of TRPM8 in human reactive lymphoid tissues and mature B-cell neoplasms

doi: 10.3892/ol.2018.9386

Figure Lengend Snippet: Frequency of the co-expression of lymphocyte markers on TRPM8-positive cells in the reactive tonsillar germinal center and SYM/marginal zone by double immunostaining.

Article Snippet: Immunohistochemistry was performed using a rabbit polyclonal anti-human TRPM8 antibody [Alomone, Jerusalem, Israel; diluted titer of 1/200; recognizes the peptide SDVD GTTYDFAHC corresponding to amino acid residues 917–929 in the 3rd extracellular loop of human TRPM8 (Accession Q7Z2W7)] ( ) as a primary antibody.

Techniques: Double Immunostaining, Standard Deviation

Journal: Oncology Letters

Article Title: Expression of TRPM8 in human reactive lymphoid tissues and mature B-cell neoplasms

doi: 10.3892/ol.2018.9386

Figure Lengend Snippet: Expression of TRPM8 in mature B-cell neoplasms.

Article Snippet: Immunohistochemistry was performed using a rabbit polyclonal anti-human TRPM8 antibody [Alomone, Jerusalem, Israel; diluted titer of 1/200; recognizes the peptide SDVD GTTYDFAHC corresponding to amino acid residues 917–929 in the 3rd extracellular loop of human TRPM8 (Accession Q7Z2W7)] ( ) as a primary antibody.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Expression of TRPM8 in human reactive lymphoid tissues and mature B-cell neoplasms

doi: 10.3892/ol.2018.9386

Figure Lengend Snippet: Relationship between IPI and TRPM8 scores. A close relationship was found between both scores ( r =−0.598855, P=0.0135). TRPM8, transient receptor potential melastatin 8; IPI, international prognostic index.

Article Snippet: Immunohistochemistry was performed using a rabbit polyclonal anti-human TRPM8 antibody [Alomone, Jerusalem, Israel; diluted titer of 1/200; recognizes the peptide SDVD GTTYDFAHC corresponding to amino acid residues 917–929 in the 3rd extracellular loop of human TRPM8 (Accession Q7Z2W7)] ( ) as a primary antibody.

Techniques:

Journal: Oncology Letters

Article Title: Expression of TRPM8 in human reactive lymphoid tissues and mature B-cell neoplasms

doi: 10.3892/ol.2018.9386

Figure Lengend Snippet: RT-PCR of TRPM8 mRNA in plasma cell myeloma, follicular lymphoma and mantle cell lymphoma. (A) A positive band of 100 bp in length for TRPM8 mRNA was found in two cases of plasma cell myeloma ( M1 & M2 ) using frozen tissue specimens. (B) A positive band of 100 bp in length for TRPM8 mRNA was found in one case of plasma cell myeloma (1–5; n=5), but not in follicular lymphoma (6,7 and 8; n=3) and mantle cell lymphoma (9,10; n=2) using FFPE tissue specimens. ( M ) Molecular markers, ( N ) negative control, ( A ) well-differentiated prostatic carcinoma as a positive control. TRPM8, transient receptor potential melastatin 8; RT-PCR, reverse transcription polymerase chain reaction.

Article Snippet: Immunohistochemistry was performed using a rabbit polyclonal anti-human TRPM8 antibody [Alomone, Jerusalem, Israel; diluted titer of 1/200; recognizes the peptide SDVD GTTYDFAHC corresponding to amino acid residues 917–929 in the 3rd extracellular loop of human TRPM8 (Accession Q7Z2W7)] ( ) as a primary antibody.

Techniques: Reverse Transcription Polymerase Chain Reaction, Negative Control, Positive Control

Journal: Oncology Letters

Article Title: Expression of TRPM8 in human reactive lymphoid tissues and mature B-cell neoplasms

doi: 10.3892/ol.2018.9386

Figure Lengend Snippet: Scheme showing the relationship between normal B-cell differentiation ( left ) and markers including TRPM8 ( right ). Activated B cells, pre-plasmablasts, and plasmablasts in the germinal center are CD20 + CD38 ± CD79a + , CD20 ± CD38 ± CD79a + IRF4/MUM-1 + , and CD38 + CD79a + TACI + IRF4/MUM-1 + , respectively. Further differentiated early and late plasma cells in the extrafollicular area are CD38 + CD79a + CD138 + TACI + IRF4/MUM-1 + and CD38 + CD79a + CD138 + TACI + , respectively. The present study demonstrated that TRPM8 is expressed on cells differentiating from pre-plasmablasts to late plasma cells. TACI (+): 10–30%, (++); 30–50%, (+++); >50% of the co-expression ratio of lymphocyte markers on TRPM8 + cells. Dotted arrows: Often positive. TRPM8, transient receptor potential melastatin 8; CD, cluster of differentiation; MUM, multiple myeloma oncogene; TACI, transmembrane activator and calcium-signal modulating cyclophilin ligand interactor.

Article Snippet: Immunohistochemistry was performed using a rabbit polyclonal anti-human TRPM8 antibody [Alomone, Jerusalem, Israel; diluted titer of 1/200; recognizes the peptide SDVD GTTYDFAHC corresponding to amino acid residues 917–929 in the 3rd extracellular loop of human TRPM8 (Accession Q7Z2W7)] ( ) as a primary antibody.

Techniques: Cell Differentiation, Expressing