Journal: Nutrients

Article Title: Long-Term Consumption of a Sugar-Sweetened Soft Drink in Combination with a Western-Type Diet Is Associated with Morphological and Molecular Changes of Taste Markers Independent of Body Weight Development in Mice

doi: 10.3390/nu14030594

Figure Lengend Snippet: Transcriptome analysis of 59 selected genes associated with oral chemosensation, displayed as a heatmap showing mean fold changes in gene expression of the SSB-fed diet groups in relation to the respective water-fed group (=1) in the form of a color code. The gene expression was analyzed using one customized cDNA microarray per group from pooled RNA samples of the CV from mice that received either a standard diet (chow, n = 10–11) or Western-type diet (WD, n = 7–8) with water (Water) or a sugar-sweetened beverage (SSB) as a drink for 24 weeks.

Article Snippet: The isolated RNA samples per mouse were reverse transcribed to cDNA using the LunaScript RT Supermix Kit (New England Biolabs GmbH, Frankfurt am Main, Germany).

Techniques: Expressing, Microarray, Western Blot

Journal:

Article Title: pp32 Reduction Induces Differentiation of TSU-Pr1 Cells

doi:

Figure Lengend Snippet: Expression of IL-6 in TSU-Pr1 cells. TSU-Pr1 cells stably transfected with pp32 anti-sense express higher levels of IL-6 message as compared to parental TSU-Pr1 cells and vector-only control by RT-PCR analysis, which validates the cDNA microarray analysis (see Figure 6).

Article Snippet: Microarray Analysis of TSU-Pr1 Cell Lines This procedure was performed at The Johns Hopkins University Oncology Microarray facility by using a human glass 12K cDNA chip.

Techniques: Expressing, Stable Transfection, Transfection, Plasmid Preparation, Control, Reverse Transcription Polymerase Chain Reaction, Microarray

Journal: Cancer cell

Article Title: RasGRP3 Mediates MAPK Pathway Activation in GNAQ Mutant Uveal Melanoma

doi: 10.1016/j.ccell.2017.04.002

Figure Lengend Snippet: (A) GNAQQ209L transfected alone or combined with PKC δ or ε into 293FT cells increased Ras-GTP levels. 293FT cells were transfected with indicated cDNA plasmids for 24 hr. Ras-GTP pull-down was performed and detected by western blot with a pan-Ras antibody. TPA stimulation was used a positive control. Expression levels of GNAQQ209L were monitored with Glu-Glu (EE) tag. PKC δ and ε were detected via HA tags.

Article Snippet: cDNA: HA-PKC δ(kinase dead) , addgene , 16389.

Techniques: Transfection, Western Blot, Positive Control, Expressing

Journal: Cancer cell

Article Title: RasGRP3 Mediates MAPK Pathway Activation in GNAQ Mutant Uveal Melanoma

doi: 10.1016/j.ccell.2017.04.002

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: cDNA: HA-PKC δ(kinase dead) , addgene , 16389.

Techniques: Recombinant, Activation Assay, Microarray, Fractionation, Expressing, Cell Culture

Journal: Cancer cell

Article Title: RasGRP3 Mediates MAPK Pathway Activation in GNAQ Mutant Uveal Melanoma

doi: 10.1016/j.ccell.2017.04.002

Figure Lengend Snippet: (A) GNAQQ209L transfected alone or combined with PKC δ or ε into 293FT cells increased Ras-GTP levels. 293FT cells were transfected with indicated cDNA plasmids for 24 hr. Ras-GTP pull-down was performed and detected by western blot with a pan-Ras antibody. TPA stimulation was used a positive control. Expression levels of GNAQQ209L were monitored with Glu-Glu (EE) tag. PKC δ and ε were detected via HA tags.

Article Snippet: cDNA: HA-PKC α (catalytic domain) , addgene , 21234.

Techniques: Transfection, Western Blot, Positive Control, Expressing

Journal: Cancer cell

Article Title: RasGRP3 Mediates MAPK Pathway Activation in GNAQ Mutant Uveal Melanoma

doi: 10.1016/j.ccell.2017.04.002

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: cDNA: HA-PKC α (catalytic domain) , addgene , 21234.

Techniques: Recombinant, Activation Assay, Microarray, Fractionation, Expressing, Cell Culture

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: (A) Representative examples of immunohistochemical staining for CAMK2N1 in each of the clinical stages of prostate cancers as indicated. (B) Quantification of CAMK2N1 relative immunostaining intensity for each clinical stage of prostate cancers. Data is shown as mean ± SEM for N as indicated in the figure in parethesis as shown. (C) CAMK2N1 mRNA determined by quantitative PCR. Comparison was made between normal and tumorous prostate samples. (D) QT-PCR analysis of CAMK2N1 expression in prostate cell lines (RWPE1, LNCaP, DU145, PC3).

Article Snippet: The CAMK2N1 human cDNA clone was purchased from OriGene Technologies and subcloned into the EcoRI/XhoI site of MSCV-IRES-GFP (Addgen) retroviral vector. pGIPZ Vector, pGIPZ-shCAMK2N1(V3LHS-315689, 5'-TCAATAACAACCCGCTTGC-3'), were purchased from Thermo Scientific.

Techniques: Immunohistochemical staining, Staining, Immunostaining, Real-time Polymerase Chain Reaction, Expressing

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: DU145 and PC3 cells stably overexpressed or knocked down CAMK2N1. Cells were analyzed for cell proliferation by (A-C) MTT, cell cycle by (D-E) FACS and oncogenic growth by (F-K) Colony-forming assay. Data is shown as mean ± SEM for N > 5 separate experiments.

Article Snippet: The CAMK2N1 human cDNA clone was purchased from OriGene Technologies and subcloned into the EcoRI/XhoI site of MSCV-IRES-GFP (Addgen) retroviral vector. pGIPZ Vector, pGIPZ-shCAMK2N1(V3LHS-315689, 5'-TCAATAACAACCCGCTTGC-3'), were purchased from Thermo Scientific.

Techniques: Stable Transfection

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: (A-J) Serial measurements were conducted every five days of DU145 and PC3 tumors stably expressing CAMK2N1, injected into nude mice. The data is shown as mean ± SEM for N>6 separate tumors for each group. (A, F) Images of tumors dissected out from the sacrificed mice. (B, G) The tumor size (mm3) versus days of post injection. (C, H) Reduction in tumor weight after resection at the end of experiment. (D-E, I-J) IHC staining detected the expression of CAMK2N1 in DU145 and PC3 tumor tissues of nude mice.

Article Snippet: The CAMK2N1 human cDNA clone was purchased from OriGene Technologies and subcloned into the EcoRI/XhoI site of MSCV-IRES-GFP (Addgen) retroviral vector. pGIPZ Vector, pGIPZ-shCAMK2N1(V3LHS-315689, 5'-TCAATAACAACCCGCTTGC-3'), were purchased from Thermo Scientific.

Techniques: Stable Transfection, Expressing, Injection, Immunohistochemistry

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: DU145 cells stably overexpressed CAMK2N1. These cells were analyzed for apoptosis by (A) Annexin V staining. Quantification of colony numbers and sizes were shown as mean ± SEM for N>5 separate experiments. (B) TUNEL staining was conducted to assess the effect of CAMK2N1 on cellular apoptosis in vivo . The percentage of apoptotic cells was increased in DU145 tumor tissues derived from nude mice. Quantification of TUNEL staining was shown as mean ± SEM for N=4 separate experiments. (C) IHC staining detected the expression of Bax, Bcl2, p21 and Ki67 in DU145 tumor tissue of nude mice. Overexpression of CAM2KN1 in DU145 and PC3 tumor tissue decreased Bcl-2, Ki67 protein expression and increased p21, Bax protein expression. Data for quantified IHC was shown as mean ± SEM for N = 4 tumors in each group.

Article Snippet: The CAMK2N1 human cDNA clone was purchased from OriGene Technologies and subcloned into the EcoRI/XhoI site of MSCV-IRES-GFP (Addgen) retroviral vector. pGIPZ Vector, pGIPZ-shCAMK2N1(V3LHS-315689, 5'-TCAATAACAACCCGCTTGC-3'), were purchased from Thermo Scientific.

Techniques: Stable Transfection, Staining, TUNEL Assay, In Vivo, Derivative Assay, Immunohistochemistry, Expressing, Over Expression

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: (A-B) Microarray gene expression analysis was conducted in DU145 cells stably overexpressing CAMK2N1 (>1.5-fold and p < 0.05). (A) Functional analysis of differentially expressed genes. Gene Ontology (GO) Biological Process (BP) terms were ranked by score (score > 1.3). (B) CAMK2N1 induced and repressed clusters of genes that were chosen from the pathways. (C-D) Expression levels of ErbB2, AKT, MEK1, ERK1/2, NFκβ, Bcl-2, BAX, and p21 were determined by Western blot in DU145 cells with stably overexpressing CAMK2N1. Similar changes as to those found in the microarray analysis were observed. Each figure represents three independent experiments. (E) DU145 tumors stably expressing CAMK2N1, and mRNA levels of ErbB2, Bcl-2, NF-κβ, AKT1, AR, p21, and Bax were determined by qRT-PCR. CAMK2N1 decreased ErbB2, BCL-2, NF-κβ, AKT1, AR mRNA levels and increased p21, Bax mRNA levels in DU145 tumor tissues. The data is shown as mean ± SEM for N=4 separate tumors for each group.

Article Snippet: The CAMK2N1 human cDNA clone was purchased from OriGene Technologies and subcloned into the EcoRI/XhoI site of MSCV-IRES-GFP (Addgen) retroviral vector. pGIPZ Vector, pGIPZ-shCAMK2N1(V3LHS-315689, 5'-TCAATAACAACCCGCTTGC-3'), were purchased from Thermo Scientific.

Techniques: Microarray, Expressing, Stable Transfection, Functional Assay, Western Blot, Quantitative RT-PCR