h19-pflag-cmv-2 expression vector (Millipore)
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Millipore
h19-pflag-cmv-2 expression vector
H19 Pflag Cmv 2 Expression Vector, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pflag-cmv-2+vector/pflag+cmv2/pmc11299361-54-10-16
Average 90 stars, based on 1 article reviews
H19 Pflag Cmv 2 Expression Vector, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/pflag-cmv-2+vector/pflag+cmv2/pmc11299361-54-10-16
Average 90 stars, based on 1 article reviews
h19-pflag-cmv-2 expression vector - by Bioz Stars,
2026-09
90/100 stars
Images
1) Product Images from "Regulatory role of lncH19 in RAC1 alternative splicing: implication for RAC1B expression in colorectal cancer"
Article Title: Regulatory role of lncH19 in RAC1 alternative splicing: implication for RAC1B expression in colorectal cancer
Journal: Journal of Experimental & Clinical Cancer Research : CR
doi: 10.1186/s13046-024-03139-z
Figure Legend Snippet: The sequence of the primers used for gene expression analysis
Techniques Used: Sequencing, Expressing
Figure Legend Snippet: The sequence of the biotin-labeled H19 probes used for RNA pull-down assays
Techniques Used: Sequencing
Figure Legend Snippet: RBFOX2 is involved in RAC1 alternative splicing in CRC cells. ( A - B - D , E - F - H ) QRT-PCR for the indicated mRNA in CRC cells, silenced for RBFOX2 with two different siRNA. Graphs show 2-ΔΔct calculated in silenced cells respect to relative controls (Normality test and t-test). ( C , F ) Western Blot and densitometric analyses for RAC1B in SW620 and HCT116 silenced for RBFOX2 and relative controls. For densitometric analysis data are represented as normalized OD. ( I - L ) qRT-PCR of the indicated genes in SW620 and HCT116 silenced for H19, the graphs represent the 2^- ΔΔ ct of the indicated calculated respect the expression in control cells. ( M ) Western Blot of RAC1b protein levels in CRC cells (SW620 and HCT116) in H19 silenced cells and relative control cells. Statistical analyses were performed using normality test and t-test, p-value is shown in th e graphs
Techniques Used: Alternative Splicing, Quantitative RT-PCR, Western Blot, Expressing, Control
Figure Legend Snippet: Overexpression of lncH19 upregulates RAC1B expression. ( A ) qRT-PCR to analyze H19 mRNA levels in CRC cells (SW620 and HCT116) transfected with lncH19 or empty vector. ( B ) qRT-PCR to analyze RAC1B mRNA levels in CRC cells (SW620 and HCT116) transfected with lncH19 or empty vector. ( C ) Western Blot for RAC1B in CRC cells (SW620 and HCT116), transfected with lncH19 or empty vector. lncH19 overexpression promotes RAC1B nuclear localization. ( D ) Representative confocal microscopy images of anti-RAC1b immunofluorescence showed nuclear localization of RAC1b in lncH19-overexpressing CRC cells, SW620 (upper panel) and HCT116 (lower panel). The analysis of the RAC1B nuclear signal is reported on the histogram. ( E ) Western blot analysis and densitometric analyses for RAC1B in nuclear protein fractions of CRC cells (SW620 and HCT116) overexpressing H19 cells and relative control cells. E-F) Overexpression of lncH19 upregulate CiclinD and c-Myc expression through RAC1B activity qRT-PCR for the indicated genes in SW620 ( F ) and HCT116 ( G ) transfected with pH19 or empty vector and treated or not with RAC1B inhibitor. Statistical analyses were performed using: normality test, one sample t-test to compare different conditions to pEmpty untreated cells, and Two-tails unpaired t-test to compare two groups *; p-value is shown in the graphs
Techniques Used: Over Expression, Expressing, Quantitative RT-PCR, Transfection, Plasmid Preparation, Western Blot, Confocal Microscopy, Immunofluorescence, Control, Activity Assay
Related Articles
Construct:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Generated:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Subcloning:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Plasmid Preparation:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Transfection:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Western Blot:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). MTT Assay:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Staining:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Expressing:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Migration:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Amplification:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Polymerase Chain Reaction:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). cDNA Library Assay:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Clone Assay:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Mutagenesis:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Immunofluorescence:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). Cell Fractionation:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). FLAG-tag:Article Title: Co-expression of phenylalanine hydroxylase variants and effects of interallelic complementation on in vitro enzyme activity and genotype-phenotype correlation. Article Snippet: Article history: Received 8 January 2016 Accepted 9 January 2016 Available online 12 January 2016 Background: In phenylketonuria (PKU) patients, the combination of two phenylalanine hydroxylase (PAH) alleles is the main determinant of residual enzyme activity in vivo and in vitro.. Inconsistencies in genotype-phenotype correlations have been observed in compound heterozygous patients and a particular combination of two PAH alleles may produce a phenotype that is different from the expected one, possibly due to interallelic complementation.. Methods: A dual eukaryotic vector systemwith two distinct PAH proteins N-terminally fused to different epitope tags was used to investigate the co-expression of PAH alleles reported in patients with inconsistent phenotypes. Article Title: Aminopeptidase A initiates tumorigenesis and enhances tumor cell stemness via TWIST1 upregulation in colorectal cancer Article Snippet: The ENPEP gene was cloned into a Article Title: PRRT2 Mutant Leads to Dysfunction of Glutamate Signaling Article Snippet: Human wild type PRRT2 full-length was cloned into the Xba I and Bam HI sites of the Article Title: O -GlcNAcylation Facilitates the Interaction between Keratin 18 and Isocitrate Dehydrogenases and Potentially Influencing Cholangiocarcinoma Progression Article Snippet: The FLAG-tagged K18 constructs were generated by in-frame subcloning the human KRT18 cDNA into the Article Title: Tenacissoside H Induces Apoptosis and Inhibits Migration of Colon Cancer Cells by Downregulating Expression of GOLPH3 Gene Article Snippet: The Human colon cancer cell line LoVo (Shanghai Institute of Cell Science, Chinese Academy of Sciences); TDH (20 mg/branch; batch number: 111913-201803; China Food and Drug Control Institute); RPMI1640 medium (Beijing Suolaibao Bioscience and Technology Co., Ltd.); pFLAG-CMV-2 vector (Sigma, St. Louis, MO, USA); cDNA reverse transcription kit (Invitrogen, Carlsbad, CA, USA); Lipofectamine TM 3000 transfection reagent (Invitrogen); Opti-MEM serum-free medium (Shanghai Tuoran Biotechnology Co., Ltd.); rabbit anti-human GOLPH3 polyclonal antibody (ab98023; Abcam, Cambridge, UK); phosphorylated ribosomal protein S6 kinase β 1 (p-p70S6K) antibody (sc-377529; Santa Cruz Biotechnology, Santa Cruz, USA); p70S6K antibody (sc-8418; Santa Cruz Biotechnology); β -catenin antibody (sc-7199; Santa Cruz Biotechnology); horseradish peroxidase-labeled goat anti-rabbit IgG antibody (A0208; Beyotime, Nanking, China); WB and IP protein lysate (Shanghai Lai Maple Biotechnology Co., Ltd.); trypsin powder, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), propidium iodide (PI) staining solution, and Article Title: Polo-like Kinase 1 (Plk1) Up-regulates Telomerase Activity by Affecting Human Telomerase Reverse Transcriptase (hTERT) Stability Article Snippet: FLAG-Plk1, FLAG-Plk1(1–330), FLAG-Plk1(330 – 603), FLAG-Plk1-TD (constitutively active form of Plk1), and FLAG-Plk1-KD (kinase-deficient form of Plk1) expression plasmids were made by cloning Plk1 or its mutated cDNAs into the Article Title: Cytoplasmic viral RNA-dependent RNA polymerase disrupts the intracellular splicing machinery by entering the nucleus and interfering with Prp8. Article Snippet: Construction of the mutant FLAG-3Dpol NLS, immunofluorescence microscopic analysis, and cellular fractionation The PCR product of EV71 3Dpol containing the wt NLS (KKKD) was cloned into the Article Title: Glycerol kinase interacts with nuclear receptor NR4A1 and regulates glucose metabolism in the liver Article Snippet: The human full-length NR4A1 gene was amplified by PCR from a human liver cDNA library and cloned into the expression vector pFlag-CMV-2 vector (MilliporeSigma, Burlington, MA, USA). |