human agtpbp1 protein  (Proteintech)

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: Top 20 significant genes coexpressed with C9orf72 on COXPRESdb.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Binding Assay, Sequencing

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: COXPRESdb search indicates coexpression of AGTPBP1 with C9orf72. The genes coexpressed with C9orf72 were identified by database search on COXPRESdb, as listed in . Interactive network of top 20 genes coexpressed with C9orf72 was visualized on Cytoscape web. ( A ) Correlation profile of gene expression between C9orf72 ( x axis: probe ID 225919_s_at) and AGTPBP1 ( y axis: probe ID 204500_s_at). ( B ) Molecular network of top 20 genes coexpressed with C9orf72. The color of nodes indicates yellow (nuclear), green (cytosol), blue (mitochondria), white (plasma membrane), and grey (ambiguous), determined by WoLF localization prediction program. The interaction between C9orf72 and AGTPBP1 is highlighted by red ellipse.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Expressing

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: Coexpression of AGTPBP1 and C9orf72 mRNA in human neural cells. The mRNA expression was studied by RT-PCR and qPCR in human neural tissues and cultured cells. The panels ( A – C ) represent RT-PCR of ( A ) AGTPBP1, ( B ) C9orf72, and ( C ) G3PDH. The lanes (1–10) indicate (1) the FC of the human CBR with inclusion of the reverse transcription (RT) step, (2) CBR without inclusion of the RT step, (3) astrocytes, (4) NP cells, (5) NTera2 teratocarcinoma-derived neurons (NTera2 N), (6) SK-N-SH neuroblastoma, (7) IMR-32 neuroblastoma, (8) U-373MG glioblastoma, (9) T98G glioblastoma, and (10) HMO6 immortalized microglia. The panels ( D, E ) represent qPCR of ( D ) AGTPBP1 and ( E ) C9orf72, standardized against the levels of G3PDH, serving as an internal control. The panel ( F ) indicates Pearson’s correlation between AGTPBP1 and C9orf72 mRNA expression levels in the set of eight human neural cells.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Derivative Assay

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: Molecular interaction of AGTPBP1 and C9orf72 in cultured cells. The full-length AGTPBP1 protein tagged with HA and the full-length C9orf72 protein tagged with Flag were coexpressed in HEK293 cells. The protein extract was processed for ( A ) IP with anti-Flag antibody followed by WB with anti-HA antibody or ( B ) IP with anti-HA antibody followed by WB with anti-C9orf72 antibody. The CP domain of AGTPBP1 tagged with Flag and the full-length C9orf72 protein tagged with Myc or EGFP were coexpressed in HEK293 cells. The protein extract was processed for ( C ) IP with anti-Myc antibody followed by WB with anti-Flag antibody or ( D ) IP with anti-Flag antibody followed by WB with anti-GFP antibody. The protein extract of SK-N-SH cells was processed for ( E ) IP with anti-C9orf72 antibody followed by WB with anti-AGTPBP1 antibody. The lanes represent (1) input control, (2) IP with the specific antibody, and (3) IP with normal IgG.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Cell Culture

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: Coexpression of AGTPBP1 and C9orf72 in cultured cells and human brains. The vectors of EGFP-tagged AGTPBP1 and RFP-tagged C9orf72 were cotransfected in NSC-34 motor neurons. Coexpression of AGTPBP1 and C9orf72 was studied in the CA1 hippocampal region of human brains derived from normal control (NC) subjects by double-labeling IHC. The panels ( A – I ) represent ( A, D ) (A, D) NSC-34, AGTPBP1 (green), ( B, E ) NSC-34, C9orf72 (red), ( C, F ) merge of AGTPBP1 and C9orf72 labeled with DAPI, ( G ) CA1, AGTPBP1 (red), ( H ) CA1, C9orf72 (green), and ( I ) merge of AGTPBP1 and C9orf72 labeled with DAPI.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Cell Culture, Derivative Assay, Labeling

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: IHC of AGTPBP1 and C9orf72 expression in human brains. The immunoreactivity for AGTPBP1 or C9orf72 was studied in the hippocampus CA4 region and the frontal cortex (FC) of NC and AD brains by IHC. The panels (A – D) indicate (A) NC, CA4, AGTPBP1, (B) NC, CA4, C9orf72, (C) NC, FC, AGTPBP1, and (D) AD, FC, AGTPBP1. Scale bar = 200 μm.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Expressing

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: AGTPBP1 and C9orf72 mRNA expression in human brains. The mRNA expression levels were studied by qPCR in human brain tissues derived from a reference of the human FC (REF), four NC, six ALS, four PD, and seven AD cases listed in . The expression levels were standardized against those of G3PDH, serving as an internal control: ( A ) AGTPBP1, ( B ) C9orf72, and ( C ) comparison between AD and non-AD groups. The panels indicate (left) AGTPBP1 and (right) C9orf72. ( D ) Pearson’s correlation between AGTPBP1 and C9orf72 mRNA expression levels.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Expressing, Derivative Assay

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: AGTPBP1 and C9orf72 protein expression in human brains. The protein expression levels were studied by WB in human brain tissues derived from four NC, six ALS, four PD, and seven AD cases listed in . The expression levels were standardized against those of ACTB, serving as an internal control. ( A ) WB of (a) AGTPBP1, (b) C9orf72, (c) ACTB, and (d) HSP60. ( B ) Comparison between AD and non-AD groups. The panels indicate (left) AGTPBP1 and (right) C9orf72. ( C ) Pearson’s correlation between AGTPBP1 and C9orf72 protein expression levels.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Expressing, Derivative Assay

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: Knockdown of AGTPBP1 in SK-N-SH cells. The siRNA product directed to AGTPBP1 (SI) or a negative control RNA (NEG) was introduced in SK-N-SH cells by using Lipofectamine (LPF) RNAiMax reagent, followed by processing for qPCR and WB analysis. The panels ( A , B ) represent qPCR of ( A ) AGTPBP1 and ( B ) C9orf72 standardized against the levels of G3PDH mRNA, while the panels ( C – E ) indicate WB of ( C ) AGTPBP1, ( D ) C9orf72, and ( E ) ACTB, serving as an internal control. The lanes (1–3) represent the protein extract isolated from the cells treated with (1) LPF alone, (2) NEG, or (3) SI.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Negative Control, Isolation

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: Overexpression of POU2F1 in SK-N-SH cells. The POU2F1 expression vector or the control vector expressing LacZ was transfected in SK-N-SH cells by using Lipofectamine (LPF) 2000 reagent, followed by processing for qPCR and WB analysis. The panels ( A, B ) represent qPCR of ( A ) AGTPBP1 and ( B ) C9orf72 standardized against the levels of G3PDH mRNA, while the panels ( C, D ) indicate WB of ( C ) POU2F1 and ( D ) HSP60, serving as an internal control. The lanes (1–3) represent the protein extract isolated from the cells treated with (1) LPF alone, or the cells transfected with the expression vector of (2) POU2F1 or (3) LacZ.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Over Expression, Expressing, Plasmid Preparation, Transfection, Isolation

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: Top 20 significant genes coexpressed with C9orf72 on COXPRESdb.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Binding Assay, Sequencing, Glycoproteomics

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: COXPRESdb search indicates coexpression of AGTPBP1 with C9orf72. The genes coexpressed with C9orf72 were identified by database search on COXPRESdb, as listed in . Interactive network of top 20 genes coexpressed with C9orf72 was visualized on Cytoscape web. ( A ) Correlation profile of gene expression between C9orf72 ( x axis: probe ID 225919_s_at) and AGTPBP1 ( y axis: probe ID 204500_s_at). ( B ) Molecular network of top 20 genes coexpressed with C9orf72. The color of nodes indicates yellow (nuclear), green (cytosol), blue (mitochondria), white (plasma membrane), and grey (ambiguous), determined by WoLF localization prediction program. The interaction between C9orf72 and AGTPBP1 is highlighted by red ellipse.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Gene Expression, Clinical Proteomics, Membrane

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: Coexpression of AGTPBP1 and C9orf72 mRNA in human neural cells. The mRNA expression was studied by RT-PCR and qPCR in human neural tissues and cultured cells. The panels ( A – C ) represent RT-PCR of ( A ) AGTPBP1, ( B ) C9orf72, and ( C ) G3PDH. The lanes (1–10) indicate (1) the FC of the human CBR with inclusion of the reverse transcription (RT) step, (2) CBR without inclusion of the RT step, (3) astrocytes, (4) NP cells, (5) NTera2 teratocarcinoma-derived neurons (NTera2 N), (6) SK-N-SH neuroblastoma, (7) IMR-32 neuroblastoma, (8) U-373MG glioblastoma, (9) T98G glioblastoma, and (10) HMO6 immortalized microglia. The panels ( D, E ) represent qPCR of ( D ) AGTPBP1 and ( E ) C9orf72, standardized against the levels of G3PDH, serving as an internal control. The panel ( F ) indicates Pearson’s correlation between AGTPBP1 and C9orf72 mRNA expression levels in the set of eight human neural cells.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Cell Culture, Reverse Transcription, Derivative Assay, Control

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: Molecular interaction of AGTPBP1 and C9orf72 in cultured cells. The full-length AGTPBP1 protein tagged with HA and the full-length C9orf72 protein tagged with Flag were coexpressed in HEK293 cells. The protein extract was processed for ( A ) IP with anti-Flag antibody followed by WB with anti-HA antibody or ( B ) IP with anti-HA antibody followed by WB with anti-C9orf72 antibody. The CP domain of AGTPBP1 tagged with Flag and the full-length C9orf72 protein tagged with Myc or EGFP were coexpressed in HEK293 cells. The protein extract was processed for ( C ) IP with anti-Myc antibody followed by WB with anti-Flag antibody or ( D ) IP with anti-Flag antibody followed by WB with anti-GFP antibody. The protein extract of SK-N-SH cells was processed for ( E ) IP with anti-C9orf72 antibody followed by WB with anti-AGTPBP1 antibody. The lanes represent (1) input control, (2) IP with the specific antibody, and (3) IP with normal IgG.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Cell Culture, Control

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: Coexpression of AGTPBP1 and C9orf72 in cultured cells and human brains. The vectors of EGFP-tagged AGTPBP1 and RFP-tagged C9orf72 were cotransfected in NSC-34 motor neurons. Coexpression of AGTPBP1 and C9orf72 was studied in the CA1 hippocampal region of human brains derived from normal control (NC) subjects by double-labeling IHC. The panels ( A – I ) represent ( A, D ) (A, D) NSC-34, AGTPBP1 (green), ( B, E ) NSC-34, C9orf72 (red), ( C, F ) merge of AGTPBP1 and C9orf72 labeled with DAPI, ( G ) CA1, AGTPBP1 (red), ( H ) CA1, C9orf72 (green), and ( I ) merge of AGTPBP1 and C9orf72 labeled with DAPI.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Cell Culture, Derivative Assay, Control, Labeling

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: IHC of AGTPBP1 and C9orf72 expression in human brains. The immunoreactivity for AGTPBP1 or C9orf72 was studied in the hippocampus CA4 region and the frontal cortex (FC) of NC and AD brains by IHC. The panels (A – D) indicate (A) NC, CA4, AGTPBP1, (B) NC, CA4, C9orf72, (C) NC, FC, AGTPBP1, and (D) AD, FC, AGTPBP1. Scale bar = 200 μm.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Expressing

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: AGTPBP1 and C9orf72 mRNA expression in human brains. The mRNA expression levels were studied by qPCR in human brain tissues derived from a reference of the human FC (REF), four NC, six ALS, four PD, and seven AD cases listed in . The expression levels were standardized against those of G3PDH, serving as an internal control: ( A ) AGTPBP1, ( B ) C9orf72, and ( C ) comparison between AD and non-AD groups. The panels indicate (left) AGTPBP1 and (right) C9orf72. ( D ) Pearson’s correlation between AGTPBP1 and C9orf72 mRNA expression levels.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Expressing, Derivative Assay, Control, Comparison

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: AGTPBP1 and C9orf72 protein expression in human brains. The protein expression levels were studied by WB in human brain tissues derived from four NC, six ALS, four PD, and seven AD cases listed in . The expression levels were standardized against those of ACTB, serving as an internal control. ( A ) WB of (a) AGTPBP1, (b) C9orf72, (c) ACTB, and (d) HSP60. ( B ) Comparison between AD and non-AD groups. The panels indicate (left) AGTPBP1 and (right) C9orf72. ( C ) Pearson’s correlation between AGTPBP1 and C9orf72 protein expression levels.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Expressing, Derivative Assay, Control, Comparison

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: Knockdown of AGTPBP1 in SK-N-SH cells. The siRNA product directed to AGTPBP1 (SI) or a negative control RNA (NEG) was introduced in SK-N-SH cells by using Lipofectamine (LPF) RNAiMax reagent, followed by processing for qPCR and WB analysis. The panels ( A , B ) represent qPCR of ( A ) AGTPBP1 and ( B ) C9orf72 standardized against the levels of G3PDH mRNA, while the panels ( C – E ) indicate WB of ( C ) AGTPBP1, ( D ) C9orf72, and ( E ) ACTB, serving as an internal control. The lanes (1–3) represent the protein extract isolated from the cells treated with (1) LPF alone, (2) NEG, or (3) SI.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Knockdown, Negative Control, Control, Isolation

Journal: Journal of Central Nervous System Disease

Article Title: Bioinformatics Data Mining Approach Suggests Coexpression of AGTPBP1 with an ALS-linked Gene C9orf72

doi: 10.4137/JCNSD.S24317

Figure Lengend Snippet: Overexpression of POU2F1 in SK-N-SH cells. The POU2F1 expression vector or the control vector expressing LacZ was transfected in SK-N-SH cells by using Lipofectamine (LPF) 2000 reagent, followed by processing for qPCR and WB analysis. The panels ( A, B ) represent qPCR of ( A ) AGTPBP1 and ( B ) C9orf72 standardized against the levels of G3PDH mRNA, while the panels ( C, D ) indicate WB of ( C ) POU2F1 and ( D ) HSP60, serving as an internal control. The lanes (1–3) represent the protein extract isolated from the cells treated with (1) LPF alone, or the cells transfected with the expression vector of (2) POU2F1 or (3) LacZ.

Article Snippet: Then, the tissue sections were incubated at 4 °C overnight with a rabbit polyclonal anti-AGTPBP1 antibody raised against a peptide spanning amino acid residues 837–1186 of the human AGTPBP1 protein (14067–1-AP; Proteintech), a rabbit polyclonal anti-human C9orf72 antibody raised against a peptide spanning amino acid residues 165–215 of the human C9orf72 protein (sc-138763; Santa Cruz Biotechnology), whose specificity was validated previously, or a mouse monoclonal anti-AGTPBP1 antibody raised against a recombinant protein fragment spanning amino acid residues 368–753 of the human AGTPBP1 (9A3; LifeSpan Biosciences).

Techniques: Over Expression, Expressing, Plasmid Preparation, Control, Transfection, Isolation