Journal: Alzheimer's Research & Therapy

Article Title: TMEM106B expression is reduced in Alzheimer’s disease brains

doi: 10.1186/alzrt247

Figure Lengend Snippet: Multiple sequence alignment of TMEM106B protein. Multiple amino acid sequence alignment was performed by importing the corresponding amino acid sequences into CLC Free Workbench (CLC Bio/Qiagen, Aarhus, Denmark). (a) Multiple amino acid sequence alignment of TMEM106B orthologs derived from Homo sapiens , Pan troglodytes , Canis lupus familiaris , Bos Taurus , Mus musclus , Rattus norvegicus , Gallus gallus , Danio rerio , and Xenopus laevis . (b) Multiple amino acid sequence alignment of the human TMEM106A, TMEM106B, and TMEM106C proteins.

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: Sequencing, Derivative Assay

Journal: Alzheimer's Research & Therapy

Article Title: TMEM106B expression is reduced in Alzheimer’s disease brains

doi: 10.1186/alzrt247

Figure Lengend Snippet: Universal expression of TMEM106B mRNAs in human neural cells. mRNA expression was studied by reverse transcriptase (RT)-polymerase chain reaction (PCR) in human tissues and cultured cells. (a) TMEM106A, (b) TMEM106B, (c) TMEM106C, (d) progranulin (PGRN), and (e) G3PDH, a housekeeping gene for a positive control. The lanes indicate (1) the frontal cortex of the human cerebrum (CBR) with inclusion of the RT step, (2) CBR without inclusion of the RT step, (3) astrocytes (AS), (4) neuronal progenitor (NP) cells, (5) NTera2 teratocarcinoma-derived neurons, (6) SK-N-SH neuroblastoma, (7) IMR-32 neuroblastoma, (8) U-373MG glioblastoma, (9) T98G glioblastoma, and (10) HMO6 microglia. TMEM106A, TMEM106B, TMEM106C, and PGRN were amplified for 35 cycles, while G3PDH was amplified for 28 cycles.

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: Expressing, Polymerase Chain Reaction, Cell Culture, Positive Control, Derivative Assay, Amplification

Journal: Alzheimer's Research & Therapy

Article Title: TMEM106B expression is reduced in Alzheimer’s disease brains

doi: 10.1186/alzrt247

Figure Lengend Snippet: Reduced expression of TMEM106B mRNA in Alzheimer’s disease brains. TMEM106B and progranulin (PGRN) mRNA expression levels were studied by quantitative reverse transcriptase-polymerase chain reaction (qPCR) in human brain tissues derived from a reference of the human frontal cortex (REF), four non-neurological control cases (NC), six amyotrophic lateral sclerosis (ALS) cases, four Parkinson’s disease (PD) cases, and seven Alzheimer’s disease (AD) cases. The expression levels were standardized against those of G3PDH. (a) TMEM106B mRNA expression. (b) PGRN mRNA expression. (c) Difference in TMEM106B levels between AD and non-AD cases. * P = 0.0035 by Student’s t test. (d) Difference in PGRN levels between AD and non-AD cases. ** P = 0.0027 by Student’s t test. (e) Pearson’s correlation between TMEM106B and PGRN mRNA levels. Pearson’s correlation coefficient indicates −0.555 ( P = 0.0090).

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: Expressing, Polymerase Chain Reaction, Derivative Assay

Journal: Alzheimer's Research & Therapy

Article Title: TMEM106B expression is reduced in Alzheimer’s disease brains

doi: 10.1186/alzrt247

Figure Lengend Snippet: Positive correlation between TMEM106B and neurofilament, heavy polypeptide mRNA levels. Neurofilament, heavy polypeptide (NFH), glial fibrillary acidic protein (GFAP), and RNA-binding protein, fox-1 homolog ( Caenorhabditis elegans )-3 (RBFOX3, NEUN) mRNA expression levels were studied by quantitative reverse transcriptase-polymerase chain reaction (qPCR) in human brain tissues derived from a reference of the human frontal cortex (REF), four non-neurological causes (NC) cases, six amyotrophic lateral sclerosis (ALS) cases, four Parkinson’s disease PD cases, and seven AD cases. The expression levels were standardized against those of G3PDH. (a) NFH expression. (b) GFAP expression. (c) NEUN expression. (d) Pearson’s correlation between TMEM106B and NFH mRNA levels. Pearson’s correlation coefficient indicates 0.496 ( P = 0.0221).

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: RNA Binding Assay, Expressing, Polymerase Chain Reaction, Derivative Assay

Journal: Alzheimer's Research & Therapy

Article Title: TMEM106B expression is reduced in Alzheimer’s disease brains

doi: 10.1186/alzrt247

Figure Lengend Snippet: Characterization of anti-TMEM106B antibody. The full-length open reading frame (ORF) cloned in the vector that expresses a fusion protein with an N-terminal Xpress tag was transiently expressed in HeLa cells. Total protein extract was processed for western blot. Lanes represent the protein of (1) untransfected cells and the cells expressing (2) TMEM106A, (3) TMEM106B, or (4) TMEM106C, and the protein of (5) human brain #1, (6) human brain #2, or (7) IMR-32 neuroblastoma cells. Immunoblots of (a, d) TMEM106B (the A303-439A antibody), (b) Xpress, and (c, e) HSP60, an internal control for protein loading.

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: Clone Assay, Plasmid Preparation, Western Blot, Expressing

Journal: Alzheimer's Research & Therapy

Article Title: TMEM106B expression is reduced in Alzheimer’s disease brains

doi: 10.1186/alzrt247

Figure Lengend Snippet: Reduced expression of TMEM106B protein in Alzheimer’s disease brains. Protein expression levels were studied by western blot in human brain tissues derived from four non-neurological causes (NC) cases, six amyotrophic lateral sclerosis (ALS) cases, four Parkinson’s disease (PD) cases, and seven Alzheimer’s disease (AD) cases. The expression levels were standardized against those of HSP60. (A) TMEM106B expression: (a) TMEM106B and (b) HSP60. (B) Progranulin (PGRN) expression: (a) PGRN and (b) HSP60. (C) Difference in TMEM106B levels between AD and non-AD cases. * P = 0.0000004 by Student’s t test. (D) Difference in PGRN levels between AD and non-AD cases. ns, non-significant ( P = 0.5304 by Student’s t test). (E) Pearson’s correlation between TMEM106B and PGRN protein levels. Pearson’s correlation coefficient indicates −0.242 ( P = 0.2912).

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: Expressing, Western Blot, Derivative Assay

Journal: Alzheimer's Research & Therapy

Article Title: TMEM106B expression is reduced in Alzheimer’s disease brains

doi: 10.1186/alzrt247

Figure Lengend Snippet: TMEM106B immunoreactivity in non-Alzheimer’s disease brains. Expression of TMEM106 immunoreactivity was studied in 13 non-Alzheimer’s disease brains presented in Table by immunohistochemistry using the A303-439A antibody. (a) Non-neurological causes (NC), the frontal cortex, cytoplasmic staining of cortical neurons; (b) amyotrophic lateral sclerosis (ALS), the frontal cortex, cytoplasmic staining of cortical neurons; (c) NC, the hippocampal CA1 region, cytoplasmic staining of pyramidal neurons; (d) ALS, the hippocampal CA1 region, cytoplasmic staining of pyramidal neurons; (e) NC, the hippocampal CA1 region, intense staining of small nodular structures accumulated in the perinuclear region of pyramidal neurons; (f) NC, the frontal white matter, cytoplasmic staining of oligodendrocytes, reactive astrocytes, and microglia.

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Alzheimer's Research & Therapy

Article Title: TMEM106B expression is reduced in Alzheimer’s disease brains

doi: 10.1186/alzrt247

Figure Lengend Snippet: TMEM106B and PGRN immunoreactivities in Alzheimer’s disease brains. Expression of TMEM106 and progranulin (PGRN) immunoreactivities was studied in six Alzheimer’s disease brains presented in Table by immunohistochemistry using the A303-439A antibody. (a) TMEM106B, the frontal cortex, moderate neuronal cytoplasmic staining and faint senile plaque staining; (b) PGRN, same region as (a) , moderate senile plaque staining and diffuse neuropil staining; (c) TMEM106B, the hippocampal CA1 region, intense neuronal and astroglial cytoplasmic staining; (d) PGRN, same region as (c) , intense perivascular neuropil staining; (e) TMEM106B, the hippocampal CA1 region, no staining of senile plaques and neurofibrillary tangles; (f) PGRN, same region as (e) , moderate staining of numerous senile plaques and neurofibrillary tangles.

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Alzheimer's research & therapy

Article Title: TMEM106B expression is reduced in Alzheimer's disease brains.

doi: 10.1186/alzrt247

Figure Lengend Snippet: Figure 1 Multiple sequence alignment of TMEM106B protein. Multiple amino acid sequence alignment was performed by importing the corresponding amino acid sequences into CLC Free Workbench (CLC Bio/Qiagen, Aarhus, Denmark). (a) Multiple amino acid sequence alignment of TMEM106B orthologs derived from Homo sapiens, Pan troglodytes, Canis lupus familiaris, Bos Taurus, Mus musclus, Rattus norvegicus, Gallus gallus, Danio rerio, and Xenopus laevis. (b) Multiple amino acid sequence alignment of the human TMEM106A, TMEM106B, and TMEM106C proteins.

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: Sequencing, Derivative Assay

Journal: Alzheimer's research & therapy

Article Title: TMEM106B expression is reduced in Alzheimer's disease brains.

doi: 10.1186/alzrt247

Figure Lengend Snippet: Figure 2 Universal expression of TMEM106B mRNAs in human neural cells. mRNA expression was studied by reverse transcriptase (RT)-polymerase chain reaction (PCR) in human tissues and cultured cells. (a) TMEM106A, (b) TMEM106B, (c) TMEM106C, (d) progranulin (PGRN), and (e) G3PDH, a housekeeping gene for a positive control. The lanes indicate (1) the frontal cortex of the human cerebrum (CBR) with inclusion of the RT step, (2) CBR without inclusion of the RT step, (3) astrocytes (AS), (4) neuronal progenitor (NP) cells, (5) NTera2 teratocarcinoma-derived neurons, (6) SK-N-SH neuroblastoma, (7) IMR-32 neuroblastoma, (8) U-373MG glioblastoma, (9) T98G glioblastoma, and (10) HMO6 microglia. TMEM106A, TMEM106B, TMEM106C, and PGRN were amplified for 35 cycles, while G3PDH was amplified for 28 cycles.

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

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

Journal: Alzheimer's research & therapy

Article Title: TMEM106B expression is reduced in Alzheimer's disease brains.

doi: 10.1186/alzrt247

Figure Lengend Snippet: Figure 3 Reduced expression of TMEM106B mRNA in Alzheimer’s disease brains. TMEM106B and progranulin (PGRN) mRNA expression levels were studied by quantitative reverse transcriptase-polymerase chain reaction (qPCR) in human brain tissues derived from a reference of the human frontal cortex (REF), four non-neurological control cases (NC), six amyotrophic lateral sclerosis (ALS) cases, four Parkinson’s disease (PD) cases, and seven Alzheimer’s disease (AD) cases. The expression levels were standardized against those of G3PDH. (a) TMEM106B mRNA expression. (b) PGRN mRNA expression. (c) Difference in TMEM106B levels between AD and non-AD cases. *P = 0.0035 by Student’s t test. (d) Difference in PGRN levels between AD and non-AD cases. **P = 0.0027 by Student’s t test. (e) Pearson’s correlation between TMEM106B and PGRN mRNA levels. Pearson’s correlation coefficient indicates −0.555 (P = 0.0090).

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

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

Journal: Alzheimer's research & therapy

Article Title: TMEM106B expression is reduced in Alzheimer's disease brains.

doi: 10.1186/alzrt247

Figure Lengend Snippet: Figure 4 Positive correlation between TMEM106B and neurofilament, heavy polypeptide mRNA levels. Neurofilament, heavy polypeptide (NFH), glial fibrillary acidic protein (GFAP), and RNA-binding protein, fox-1 homolog (Caenorhabditis elegans)-3 (RBFOX3, NEUN) mRNA expression levels were studied by quantitative reverse transcriptase-polymerase chain reaction (qPCR) in human brain tissues derived from a reference of the human frontal cortex (REF), four non-neurological causes (NC) cases, six amyotrophic lateral sclerosis (ALS) cases, four Parkinson’s disease PD cases, and seven AD cases. The expression levels were standardized against those of G3PDH. (a) NFH expression. (b) GFAP expression. (c) NEUN expression. (d) Pearson’s correlation between TMEM106B and NFH mRNA levels. Pearson’s correlation coefficient indicates 0.496 (P = 0.0221).

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: RNA Binding Assay, Expressing, Reverse Transcription, Polymerase Chain Reaction, Derivative Assay

Journal: Alzheimer's research & therapy

Article Title: TMEM106B expression is reduced in Alzheimer's disease brains.

doi: 10.1186/alzrt247

Figure Lengend Snippet: Figure 5 Characterization of anti-TMEM106B antibody. The full-length open reading frame (ORF) cloned in the vector that expresses a fusion protein with an N-terminal Xpress tag was transiently expressed in HeLa cells. Total protein extract was processed for western blot. Lanes represent the protein of (1) untransfected cells and the cells expressing (2) TMEM106A, (3) TMEM106B, or (4) TMEM106C, and the protein of (5) human brain #1, (6) human brain #2, or (7) IMR-32 neuroblastoma cells. Immunoblots of (a, d) TMEM106B (the A303-439A antibody), (b) Xpress, and (c, e) HSP60, an internal control for protein loading.

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: Clone Assay, Plasmid Preparation, Western Blot, Expressing, Control

Journal: Alzheimer's research & therapy

Article Title: TMEM106B expression is reduced in Alzheimer's disease brains.

doi: 10.1186/alzrt247

Figure Lengend Snippet: Figure 6 Reduced expression of TMEM106B protein in Alzheimer’s disease brains. Protein expression levels were studied by western blot in human brain tissues derived from four non-neurological causes (NC) cases, six amyotrophic lateral sclerosis (ALS) cases, four Parkinson’s disease (PD) cases, and seven Alzheimer’s disease (AD) cases. The expression levels were standardized against those of HSP60. (A) TMEM106B expression: (a) TMEM106B and (b) HSP60. (B) Progranulin (PGRN) expression: (a) PGRN and (b) HSP60. (C) Difference in TMEM106B levels between AD and non-AD cases. *P = 0.0000004 by Student’s t test. (D) Difference in PGRN levels between AD and non-AD cases. ns, non-significant (P = 0.5304 by Student’s t test). (E) Pearson’s correlation between TMEM106B and PGRN protein levels. Pearson’s correlation coefficient indicates −0.242 (P = 0.2912).

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: Expressing, Western Blot, Derivative Assay

Journal: Alzheimer's research & therapy

Article Title: TMEM106B expression is reduced in Alzheimer's disease brains.

doi: 10.1186/alzrt247

Figure Lengend Snippet: Figure 7 TMEM106B immunoreactivity in non-Alzheimer’s disease brains. Expression of TMEM106 immunoreactivity was studied in 13 non-Alzheimer’s disease brains presented in Table 1 by immunohistochemistry using the A303-439A antibody. (a) Non-neurological causes (NC), the frontal cortex, cytoplasmic staining of cortical neurons; (b) amyotrophic lateral sclerosis (ALS), the frontal cortex, cytoplasmic staining of cortical neurons; (c) NC, the hippocampal CA1 region, cytoplasmic staining of pyramidal neurons; (d) ALS, the hippocampal CA1 region, cytoplasmic staining of pyramidal neurons; (e) NC, the hippocampal CA1 region, intense staining of small nodular structures accumulated in the perinuclear region of pyramidal neurons; (f) NC, the frontal white matter, cytoplasmic staining of oligodendrocytes, reactive astrocytes, and microglia.

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Alzheimer's research & therapy

Article Title: TMEM106B expression is reduced in Alzheimer's disease brains.

doi: 10.1186/alzrt247

Figure Lengend Snippet: Figure 8 TMEM106B and PGRN immunoreactivities in Alzheimer’s disease brains. Expression of TMEM106 and progranulin (PGRN) immunoreactivities was studied in six Alzheimer’s disease brains presented in Table 1 by immunohistochemistry using the A303-439A antibody. (a) TMEM106B, the frontal cortex, moderate neuronal cytoplasmic staining and faint senile plaque staining; (b) PGRN, same region as (a), moderate senile plaque staining and diffuse neuropil staining; (c) TMEM106B, the hippocampal CA1 region, intense neuronal and astroglial cytoplasmic staining; (d) PGRN, same region as (c), intense perivascular neuropil staining; (e) TMEM106B, the hippocampal CA1 region, no staining of senile plaques and neurofibrillary tangles; (f) PGRN, same region as (e), moderate staining of numerous senile plaques and neurofibrillary tangles.

Article Snippet: After gel electrophoresis, the protein was transferred onto nitrocellulose membranes, followed by incubation at room temperature overnight with the anti-TMEM106B antibody A303-439A, rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 101 to 200 of the human TMEM106B protein (bs-11694R; Bioss, Boston, MA, USA), rabbit polyclonal anti-TMEM106B antibody raised against the human TMEM106B-GST fusion protein (20995-1-AP; Proteintech, Chicago, IL, USA), or mouse monoclonal anti-Xpress antibody (Invitrogen).

Techniques: Expressing, Immunohistochemistry, Staining

Journal: Acta neuropathologica

Article Title: TMEM106B amyloid filaments in the Biondi bodies of ependymal cells.

doi: 10.1007/s00401-024-02807-w

Figure Lengend Snippet: Fig. 3 Immunoblotting of Biondi bodies using antibody TMEM239 and genotyping. Sarkosyl-insoluble extracts from the choroid plex- uses of cases 3 and 12–20 and extracts from the ependymal linings of the lateral ventricles from cases 3 and 12 were immunoblotted using antibody TMEM239, which labels TMEM106B inclusions specifi- cally. All extracts showed an immunoreactive band of 29 kDa, diag- nostic of the presence of TMEM106B filaments

Article Snippet: The following anti-TMEM106B antibodies were used: TMEM239 (1:500), a rabbit polyclonal antibody raised against a synthetic peptide corresponding to residues 239–250 of human TMEM106B [54]; A303-439A (1:200), a mouse monoclonal antibody specific for the N-terminal 50 amino acids of human TMEM106B (Bethyl Laboratories); TMEM263 (1:200), a rabbit polyclonal antibody raised against a synthetic peptide corresponding to residues 263–274 of human TMEM106B [3].

Techniques: Western Blot

Journal: Acta neuropathologica

Article Title: TMEM106B amyloid filaments in the Biondi bodies of ependymal cells.

doi: 10.1007/s00401-024-02807-w

Figure Lengend Snippet: Fig. 6 Immunoelectron microscopy of a Biondi body. Choroid plex- uses from case 8 and anti-TMEM106B antibody TMEM239. The cytoplasm of an ependymal cell contains a round body made of osmi- ophilic and electron-lucent areas that are surrounded by a crescent- shaped bundle of filaments (interpreted as a Biondi body) decorated by gold particles. The round body has the appearance of a secondary lysosome or a residual body. The nucleus of the cell and numerous mitochondria are seen around the inclusion. Scale bar, 1 µm

Article Snippet: The following anti-TMEM106B antibodies were used: TMEM239 (1:500), a rabbit polyclonal antibody raised against a synthetic peptide corresponding to residues 239–250 of human TMEM106B [54]; A303-439A (1:200), a mouse monoclonal antibody specific for the N-terminal 50 amino acids of human TMEM106B (Bethyl Laboratories); TMEM263 (1:200), a rabbit polyclonal antibody raised against a synthetic peptide corresponding to residues 263–274 of human TMEM106B [3].

Techniques: Immuno-Electron Microscopy

Journal: Acta neuropathologica

Article Title: TMEM106B amyloid filaments in the Biondi bodies of ependymal cells.

doi: 10.1007/s00401-024-02807-w

Figure Lengend Snippet: Fig. 8 Immunoelectron micros- copy of Biondi bodies. Choroid plexuses from cases 8 and 23 and anti-TMEM106B antibody TMEM239. a, A membrane- bound, oval-shaped inclusion is decorated by gold particles and contains round osmiophilic bod- ies, an electron-lucent area and wisps of filaments. Two bundles of filaments decorated by gold particles are seen outside the membrane-bound inclusion. Scale bar, 1 µm b, Multiple osmiophilic bodies of different sizes are intermixed with a large bundle of filaments decorated by gold particles. Scale bar, 1 µm

Article Snippet: The following anti-TMEM106B antibodies were used: TMEM239 (1:500), a rabbit polyclonal antibody raised against a synthetic peptide corresponding to residues 239–250 of human TMEM106B [54]; A303-439A (1:200), a mouse monoclonal antibody specific for the N-terminal 50 amino acids of human TMEM106B (Bethyl Laboratories); TMEM263 (1:200), a rabbit polyclonal antibody raised against a synthetic peptide corresponding to residues 263–274 of human TMEM106B [3].

Techniques: Membrane

Journal: Cell

Article Title: Homotypic fibrillization of TMEM106B across diverse neurodegenerative diseases.

doi: 10.1016/j.cell.2022.02.026

Figure Lengend Snippet: Figure 1. Overview of native TMEM106B (A) Schematic view of TMEM106B with its N-ter- minal domain (NTD), transmembrane domain (TM), and C-terminal domain (CTD). (B) A predicted structure of TMEM106B from ROBETTA (Kim et al., 2004) colored with the same scheme as (A), indicating sites at which the C-terminal domain may get cleaved. (C) Aggregation propensity mapped onto a pre- dicted structure of TMEM106B(120–254) from AlphaFold (Jumper et al., 2021) (positive values in blue indicate soluble regions and negative values in red correspond to aggregation-prone regions).

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-TMEM106B (204-253) Novus Biologicals Cat#NBP1-91311; RRID: AB_11019681; QC18333-42825 IRDye 800CW Donkey anti-Rabbit IgG Secondary Antibody LI-COR Biosciences Cat#926-32213; RRID: AB_621848; D11005-09 Mouse monoclonal anti-TMEM106B (1-46) Proteintech Cat#60333-1-Ig; RRID: AB_2881442 Rabbit polyclonal anti-TMEM106B (101-200) Bioss Cat#bs-11694R; RRID: AB_2905622 Rabbit polyclonal anti-TMEM106B (111-190) Biorbyt Cat#orb158617; RRID: AB_2905623 Rabbit polyclonal anti-TMEM106B (150-275) Proteintech Cat#20995-1-AP; RRID: AB_10694293 Rabbit polyclonal anti-TMEM106B (204-253) Sigma Aldrich Cat#SAB2106773; RRID AB_2905624 Rabbit polyclonal anti-TMEM106B (218-252) LSBio Cat#LS-C757550; RRID: AB_2905625 Mouse monoclonal anti-pTau (phosphorylated at Ser262 and Ser356) P. Seubert, Elan Pharmaceuticals; San Francisco, CA; USA 12E8 Mouse monoclonal anti-pTau (phosphorylated at Ser396 and Ser404) P. Davies, Albert Einstein College of Medicine; New York, NY; USA PHF1; RRID: AB_2315150 Mouse monoclonal anti-pTau (phosphorylated at Ser202) P. Davies, Albert Einstein College of Medicine; New York, NY; USA CP13; RRID: AB_2314223 Rabbit polyclonal anti-Tau (Human-specific) L. Petrucelli, Mayo Clinic; Jacksonville, FL; USA

Techniques:

Journal: Cell

Article Title: Homotypic fibrillization of TMEM106B across diverse neurodegenerative diseases.

doi: 10.1016/j.cell.2022.02.026

Figure Lengend Snippet: Figure 2. Cryo-EM reconstructions of TMEM106B fibrils Cross-sections (10 z-slice average) of the TMEM106B(120–254) singlet and doublet fibril unsharpened density maps from eight cases of FTLD-TDP, two cases of PSP, and one case of DLB. See also Figures S1–S3 and S6 and Tables S2 and S3.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-TMEM106B (204-253) Novus Biologicals Cat#NBP1-91311; RRID: AB_11019681; QC18333-42825 IRDye 800CW Donkey anti-Rabbit IgG Secondary Antibody LI-COR Biosciences Cat#926-32213; RRID: AB_621848; D11005-09 Mouse monoclonal anti-TMEM106B (1-46) Proteintech Cat#60333-1-Ig; RRID: AB_2881442 Rabbit polyclonal anti-TMEM106B (101-200) Bioss Cat#bs-11694R; RRID: AB_2905622 Rabbit polyclonal anti-TMEM106B (111-190) Biorbyt Cat#orb158617; RRID: AB_2905623 Rabbit polyclonal anti-TMEM106B (150-275) Proteintech Cat#20995-1-AP; RRID: AB_10694293 Rabbit polyclonal anti-TMEM106B (204-253) Sigma Aldrich Cat#SAB2106773; RRID AB_2905624 Rabbit polyclonal anti-TMEM106B (218-252) LSBio Cat#LS-C757550; RRID: AB_2905625 Mouse monoclonal anti-pTau (phosphorylated at Ser262 and Ser356) P. Seubert, Elan Pharmaceuticals; San Francisco, CA; USA 12E8 Mouse monoclonal anti-pTau (phosphorylated at Ser396 and Ser404) P. Davies, Albert Einstein College of Medicine; New York, NY; USA PHF1; RRID: AB_2315150 Mouse monoclonal anti-pTau (phosphorylated at Ser202) P. Davies, Albert Einstein College of Medicine; New York, NY; USA CP13; RRID: AB_2314223 Rabbit polyclonal anti-Tau (Human-specific) L. Petrucelli, Mayo Clinic; Jacksonville, FL; USA

Techniques: Cryo-EM Sample Prep

Journal: Cell

Article Title: Homotypic fibrillization of TMEM106B across diverse neurodegenerative diseases.

doi: 10.1016/j.cell.2022.02.026

Figure Lengend Snippet: Figure 3. Cryo-EM structure of a TMEM106B doublet fibril Cryo-EM density (mesh) and atomic model (sticks) of (A) a TMEM106B doublet fibril and (B) the interface between the two protofilaments in the TMEM106B doublet fibril mediated by a non-proteinaceous, anionic cofactor (purple mesh) that binds to the sidechains of residues K178 and R180 of each protofilament. See also Figure S5 and Table S2.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-TMEM106B (204-253) Novus Biologicals Cat#NBP1-91311; RRID: AB_11019681; QC18333-42825 IRDye 800CW Donkey anti-Rabbit IgG Secondary Antibody LI-COR Biosciences Cat#926-32213; RRID: AB_621848; D11005-09 Mouse monoclonal anti-TMEM106B (1-46) Proteintech Cat#60333-1-Ig; RRID: AB_2881442 Rabbit polyclonal anti-TMEM106B (101-200) Bioss Cat#bs-11694R; RRID: AB_2905622 Rabbit polyclonal anti-TMEM106B (111-190) Biorbyt Cat#orb158617; RRID: AB_2905623 Rabbit polyclonal anti-TMEM106B (150-275) Proteintech Cat#20995-1-AP; RRID: AB_10694293 Rabbit polyclonal anti-TMEM106B (204-253) Sigma Aldrich Cat#SAB2106773; RRID AB_2905624 Rabbit polyclonal anti-TMEM106B (218-252) LSBio Cat#LS-C757550; RRID: AB_2905625 Mouse monoclonal anti-pTau (phosphorylated at Ser262 and Ser356) P. Seubert, Elan Pharmaceuticals; San Francisco, CA; USA 12E8 Mouse monoclonal anti-pTau (phosphorylated at Ser396 and Ser404) P. Davies, Albert Einstein College of Medicine; New York, NY; USA PHF1; RRID: AB_2315150 Mouse monoclonal anti-pTau (phosphorylated at Ser202) P. Davies, Albert Einstein College of Medicine; New York, NY; USA CP13; RRID: AB_2314223 Rabbit polyclonal anti-Tau (Human-specific) L. Petrucelli, Mayo Clinic; Jacksonville, FL; USA

Techniques: Cryo-EM Sample Prep

Journal: Cell

Article Title: Homotypic fibrillization of TMEM106B across diverse neurodegenerative diseases.

doi: 10.1016/j.cell.2022.02.026

Figure Lengend Snippet: Figure 4. Cryo-EM structure of a TMEM106B protofilament highlighting the key structural features (A) Cryo-EM density (mesh) and atomic model (sticks) of a TMEM106B protofilament. (B) A disulfide bond between C214 and C253. (C) Polymorphic site T185S and glycosylated asparagine at N183. (D–F) Glycosylated N164 (D), N151 (E), and N145 (F). See also Figures S4 and S5 and Table S2.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-TMEM106B (204-253) Novus Biologicals Cat#NBP1-91311; RRID: AB_11019681; QC18333-42825 IRDye 800CW Donkey anti-Rabbit IgG Secondary Antibody LI-COR Biosciences Cat#926-32213; RRID: AB_621848; D11005-09 Mouse monoclonal anti-TMEM106B (1-46) Proteintech Cat#60333-1-Ig; RRID: AB_2881442 Rabbit polyclonal anti-TMEM106B (101-200) Bioss Cat#bs-11694R; RRID: AB_2905622 Rabbit polyclonal anti-TMEM106B (111-190) Biorbyt Cat#orb158617; RRID: AB_2905623 Rabbit polyclonal anti-TMEM106B (150-275) Proteintech Cat#20995-1-AP; RRID: AB_10694293 Rabbit polyclonal anti-TMEM106B (204-253) Sigma Aldrich Cat#SAB2106773; RRID AB_2905624 Rabbit polyclonal anti-TMEM106B (218-252) LSBio Cat#LS-C757550; RRID: AB_2905625 Mouse monoclonal anti-pTau (phosphorylated at Ser262 and Ser356) P. Seubert, Elan Pharmaceuticals; San Francisco, CA; USA 12E8 Mouse monoclonal anti-pTau (phosphorylated at Ser396 and Ser404) P. Davies, Albert Einstein College of Medicine; New York, NY; USA PHF1; RRID: AB_2315150 Mouse monoclonal anti-pTau (phosphorylated at Ser202) P. Davies, Albert Einstein College of Medicine; New York, NY; USA CP13; RRID: AB_2314223 Rabbit polyclonal anti-Tau (Human-specific) L. Petrucelli, Mayo Clinic; Jacksonville, FL; USA

Techniques: Cryo-EM Sample Prep

Journal: Cell

Article Title: Homotypic fibrillization of TMEM106B across diverse neurodegenerative diseases.

doi: 10.1016/j.cell.2022.02.026

Figure Lengend Snippet: Figure 5. Cryo-EM structure of a highly twisted TMEM106B singlet fibril and comparison of singlet fibril molecular polymorphs (A) Cryo-EM density (mesh) and atomic model (sticks) of a highly twisted TMEM106B singlet fibril. (B) Magnified view of an unknown density bound to K178 in a highly twisted TMEM106B singlet fibril. (C) Overlay of the atomic models (Ca chain shown) of the low-twist (green) and high-twist (pink) singlet fibrils. (D) Comparison of secondary structure motifs formed by the low-twist (green) singlet fibril, high-twist (pink) singlet fibril, and native protein predicted by AlphaFold (blue) with experimentally determined post-translational modifications of the highly twisted TMEM106B singlet fibril. See also Figure S5 and Table S2.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-TMEM106B (204-253) Novus Biologicals Cat#NBP1-91311; RRID: AB_11019681; QC18333-42825 IRDye 800CW Donkey anti-Rabbit IgG Secondary Antibody LI-COR Biosciences Cat#926-32213; RRID: AB_621848; D11005-09 Mouse monoclonal anti-TMEM106B (1-46) Proteintech Cat#60333-1-Ig; RRID: AB_2881442 Rabbit polyclonal anti-TMEM106B (101-200) Bioss Cat#bs-11694R; RRID: AB_2905622 Rabbit polyclonal anti-TMEM106B (111-190) Biorbyt Cat#orb158617; RRID: AB_2905623 Rabbit polyclonal anti-TMEM106B (150-275) Proteintech Cat#20995-1-AP; RRID: AB_10694293 Rabbit polyclonal anti-TMEM106B (204-253) Sigma Aldrich Cat#SAB2106773; RRID AB_2905624 Rabbit polyclonal anti-TMEM106B (218-252) LSBio Cat#LS-C757550; RRID: AB_2905625 Mouse monoclonal anti-pTau (phosphorylated at Ser262 and Ser356) P. Seubert, Elan Pharmaceuticals; San Francisco, CA; USA 12E8 Mouse monoclonal anti-pTau (phosphorylated at Ser396 and Ser404) P. Davies, Albert Einstein College of Medicine; New York, NY; USA PHF1; RRID: AB_2315150 Mouse monoclonal anti-pTau (phosphorylated at Ser202) P. Davies, Albert Einstein College of Medicine; New York, NY; USA CP13; RRID: AB_2314223 Rabbit polyclonal anti-Tau (Human-specific) L. Petrucelli, Mayo Clinic; Jacksonville, FL; USA

Techniques: Cryo-EM Sample Prep, Comparison

Journal: Cell

Article Title: Homotypic fibrillization of TMEM106B across diverse neurodegenerative diseases.

doi: 10.1016/j.cell.2022.02.026

Figure Lengend Snippet: Figure 6. Structure-based model of a possible interrelationship between TMEM106B, aggregated filaments, and lysosomes in neurodegenerative diseases Under physiological conditions, TMEM106B spans the lysosomal/endosomal membrane. Upon cleavage of the C-terminal fragment, TMEM106B(120–254) fibrils may form. It is currently unknown whether TMEM106B(120–254) fibrillizes in the lumen or whether lysosomal leakage occurs and TMEM106B(120–254) fragments aggregate into fibrils in the cytosol. Based on our set of TMEM106B fibril structures, we speculate that the aggregation of TMEM106B(120–254) into fibrils leads to lysosomal dysfunction, which promotes the accumulation of aberrantly aggregated amyloid fibrils such as those formed by TDP-43 (PDB:7PY2, green sticks), tau (PDB:7P65, blue sticks), or a-synuclein.

Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit polyclonal anti-TMEM106B (204-253) Novus Biologicals Cat#NBP1-91311; RRID: AB_11019681; QC18333-42825 IRDye 800CW Donkey anti-Rabbit IgG Secondary Antibody LI-COR Biosciences Cat#926-32213; RRID: AB_621848; D11005-09 Mouse monoclonal anti-TMEM106B (1-46) Proteintech Cat#60333-1-Ig; RRID: AB_2881442 Rabbit polyclonal anti-TMEM106B (101-200) Bioss Cat#bs-11694R; RRID: AB_2905622 Rabbit polyclonal anti-TMEM106B (111-190) Biorbyt Cat#orb158617; RRID: AB_2905623 Rabbit polyclonal anti-TMEM106B (150-275) Proteintech Cat#20995-1-AP; RRID: AB_10694293 Rabbit polyclonal anti-TMEM106B (204-253) Sigma Aldrich Cat#SAB2106773; RRID AB_2905624 Rabbit polyclonal anti-TMEM106B (218-252) LSBio Cat#LS-C757550; RRID: AB_2905625 Mouse monoclonal anti-pTau (phosphorylated at Ser262 and Ser356) P. Seubert, Elan Pharmaceuticals; San Francisco, CA; USA 12E8 Mouse monoclonal anti-pTau (phosphorylated at Ser396 and Ser404) P. Davies, Albert Einstein College of Medicine; New York, NY; USA PHF1; RRID: AB_2315150 Mouse monoclonal anti-pTau (phosphorylated at Ser202) P. Davies, Albert Einstein College of Medicine; New York, NY; USA CP13; RRID: AB_2314223 Rabbit polyclonal anti-Tau (Human-specific) L. Petrucelli, Mayo Clinic; Jacksonville, FL; USA

Techniques: Membrane

Journal: Cell

Article Title: Genetic Screens Identify Host Factors for SARS-CoV-2 and Common Cold Coronaviruses

doi: 10.1016/j.cell.2020.12.004

Figure Lengend Snippet:

Article Snippet: Rabbit Anti-TMEM106B , Atlas Antibodies , Cat#HPA058342; RRID: AB_2683684.

Techniques: Virus, Recombinant, Purification, Gel Extraction, One Step RT-PCR, Sequencing, CRISPR, Library Amplification, Cloning, Software, Imaging

Journal: Molecular Neurodegeneration

Article Title: The major TMEM106B dementia risk allele affects TMEM106B protein levels, fibril formation, and myelin lipid homeostasis in the ageing human hippocampus

doi: 10.1186/s13024-023-00650-3

Figure Lengend Snippet: TMEM106B increases with age in the human brain. ( A ) Schematic of TMEM106B protein domains showing the unique peptide sequence used to identify and quantify TMEM106B protein levels in human hippocampus samples. ( B ) MS2 extracted ion chromatogram of the identified peptide (SAYVSYDVQKR) and ( C ) annotated MS2 fragmentation spectrum corresponding to the peak with retention time (RT) 11.28 min. ( D ) TMEM106B levels as a function of age in CA1 hippocampus samples from the NSW BTRC and QLD Brain Bank, ( E,F ) TMEM106B levels as a function of age in dorsolateral prefrontal cortex samples from the ( E ) Johns Hopkins Ageing (ages 30 to 68) and ( F ) Emory ADRC Brain Bank cohorts (ages 45 to 96). ( G ) Peptide-level associations of TMEM106B with age in the Emory ADRC Brain Bank cohort using peptides that map to the C-terminus (residues 130–139) (left) and the N-terminus (residues 15–27) (right). ( H ) TMEM106B mRNA levels as a function of age from GSE30272, using age ranges 0–78 (left), 30–60 (middle), and 60–80 (right)

Article Snippet: Membranes were blocked for 1 h at RT with 5% skim milk in Tris-buffered saline containing 0.1% Tween-20 (TBST), then incubated overnight at 4 °C with primary antibodies; anti-C-terminus TMEM106B antibody (residues 204–253, Novus Biologicals, #NBP1-91311) diluted 1:300 in TBST with 2% skim milk or anti-N-terminus TMEM106B antibody (residues 1–50, Bethyl Laboratories, #A303-439 A) diluted 1:500 in 5% bovine serum albumin (Sigma Aldrich #A7906).

Techniques: Sequencing

Journal: Molecular Neurodegeneration

Article Title: The major TMEM106B dementia risk allele affects TMEM106B protein levels, fibril formation, and myelin lipid homeostasis in the ageing human hippocampus

doi: 10.1186/s13024-023-00650-3

Figure Lengend Snippet: Hippocampal TMEM106B levels increase in carriers of the rs1990622-A risk allele. ( A ) (Top) Gene map of the TMEM106B locus (chromosome 7p21) with annotations of key TMEM106B SNP loci associated with neurodegenerative diseases (FTD: frontotemporal dementia; ALS: amyotrophic lateral sclerosis; AD: Alzheimer’s disease; PD: Parkinson’s disease; CTE: chronic traumatic encephalopathy; HS-ageing: hippocampal sclerosis with ageing; LATE: limbic-predominant age-related TDP-43 encephalopathy). Exons (E1-E9) are denoted in red, introns in white, and non-coding regions as a line. (Bottom) Linkage disequilibrium between rs1990622 and all SNPs located ± 500,000 bp from the TMEM106B gene as measured by the squared correlation coefficient (r 2 , where r 2 = 1 is perfect linkage, depicted in red oval). Dementia risk-associated SNPs depicted in the gene map are labelled on the linkage plot. ( B ) TMEM106B protein levels as a function of age and rs1990622 genotype. P values refer to the interaction between rs1990622 genotype and age in multiple regression adjusted for PMI and sex. ( C ) TMEM106B levels as a function of rs1990622 genotype after adjusting for age, PMI, and sex (ANOVA, F = 2.7, P = 0.07). ( D ) Western blots for fibrillar TMEM106B in the sarkosyl-insoluble fraction using an anti-C-terminus antibody. Samples were from individuals aged 67–78 (blue) and 81–95 (red) that are homozygous for the rs1990622 G/G allele or carriers of the risk allele (A + ). Braak stage of each individual is shown beneath the blot. Two sarkosyl-soluble samples were loaded onto the last two lanes of each blot to assess enrichment of TMEM106B fibrils (Sol LC). ( E ) Densitometry quantification of the 31 kDa bands observed in ( D )

Article Snippet: Membranes were blocked for 1 h at RT with 5% skim milk in Tris-buffered saline containing 0.1% Tween-20 (TBST), then incubated overnight at 4 °C with primary antibodies; anti-C-terminus TMEM106B antibody (residues 204–253, Novus Biologicals, #NBP1-91311) diluted 1:300 in TBST with 2% skim milk or anti-N-terminus TMEM106B antibody (residues 1–50, Bethyl Laboratories, #A303-439 A) diluted 1:500 in 5% bovine serum albumin (Sigma Aldrich #A7906).

Techniques: Western Blot

Journal: Molecular Neurodegeneration

Article Title: The major TMEM106B dementia risk allele affects TMEM106B protein levels, fibril formation, and myelin lipid homeostasis in the ageing human hippocampus

doi: 10.1186/s13024-023-00650-3

Figure Lengend Snippet: TMEM106B levels are correlated with proteins involved in myelination and brain ageing. ( A ) Proteins correlated with TMEM106B in multiple regression adjusted for PMI, sex, and age. Proteins significant at Q < 0.05 are boxed and labelled. ( B ) Protein set enrichment map of selected significantly enriched categories from GSEA using curated protein sets from the molecular signatures database (MSIGDB, C2). Each node represents a significantly enriched protein set at Q < 0.05 that is negatively (blue) or positively (red) associated with TMEM106B levels. Protein sets with overlapping membership are connected by edges, where the thickness indicates the number of overlapping proteins. ( C ) Protein membership of specific nodes shown in ( B ). Nodes have been numbered to identify their position in the protein set enrichment map ( B ). Proteins that were significantly correlated with TMEM106B levels at Q < 0.05 are boxed

Article Snippet: Membranes were blocked for 1 h at RT with 5% skim milk in Tris-buffered saline containing 0.1% Tween-20 (TBST), then incubated overnight at 4 °C with primary antibodies; anti-C-terminus TMEM106B antibody (residues 204–253, Novus Biologicals, #NBP1-91311) diluted 1:300 in TBST with 2% skim milk or anti-N-terminus TMEM106B antibody (residues 1–50, Bethyl Laboratories, #A303-439 A) diluted 1:500 in 5% bovine serum albumin (Sigma Aldrich #A7906).

Techniques:

Journal: Molecular Neurodegeneration

Article Title: The major TMEM106B dementia risk allele affects TMEM106B protein levels, fibril formation, and myelin lipid homeostasis in the ageing human hippocampus

doi: 10.1186/s13024-023-00650-3

Figure Lengend Snippet: TMEM106B rs1990622 genotype affects the hippocampal lipidome. ( A-B ) Heatmap of lipids that were significantly affected by rs1990622 genotype (N G/G =9, N A/G =20, N A/A =15), in individuals with APOE ε3/ε3 genotype. Each row represents a lipid that was significantly more abundant ( A ) or less abundant ( B ) in rs1990622-G/G individuals by ANOVA at Q < 0.05. ( C-D ) Top five lipid species that are more abundant ( C ) and less abundant ( D ) in rs1990622-G/G individuals ranked by P value. Lipids values are expressed as a molar % of total lipid. Cer: ceramide; DG: diacylglycerol; HexCer: hexosylceramide; Hex2Cer: dihexosylceramide; LPA: lysophosphatidic acid; LPC: lysophosphatidylcholine; PA: phosphatidic acid; PC: phosphatidylcholine; PE: phosphatidylethanolamine; PEp: phosphatidylethanolamine plasmalogen; PG: phosphatidylglycerol; PI: phosphatidylinositol; PS: phosphatidylserine; SM: sphingomyelin; ST: sulfatide; TG: triglyceride; CE: cholesterol ester

Article Snippet: Membranes were blocked for 1 h at RT with 5% skim milk in Tris-buffered saline containing 0.1% Tween-20 (TBST), then incubated overnight at 4 °C with primary antibodies; anti-C-terminus TMEM106B antibody (residues 204–253, Novus Biologicals, #NBP1-91311) diluted 1:300 in TBST with 2% skim milk or anti-N-terminus TMEM106B antibody (residues 1–50, Bethyl Laboratories, #A303-439 A) diluted 1:500 in 5% bovine serum albumin (Sigma Aldrich #A7906).

Techniques: