rabbit polyclonal anti tmem106b antibody  (Bioss)

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: 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: 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: 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: 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: 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: The sections were then incubated at 4°C overnight with a rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 1 to 50 of the human TMEM106B protein at a concentration of 0.1 μg/ml (A303-439A; Bethyl Laboratories, Montgomery, TX, USA), a rabbit monoclonal anti-PGRN antibody raised against a synthetic peptide corresponding to the residues in the human PGRN protein at a dilution of 1:1,000 (EPR3781; Abcam, Cambridge, UK), or a mouse monoclonal anti-pS409/410 TDP-43 antibody raised against a phosphopeptide composed of CMDSKpSpSGWGM at a dilution of 1:500 (TIP-PTD-M01; Cosmo Bio, Tokyo, Japan).

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: The sections were then incubated at 4°C overnight with a rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 1 to 50 of the human TMEM106B protein at a concentration of 0.1 μg/ml (A303-439A; Bethyl Laboratories, Montgomery, TX, USA), a rabbit monoclonal anti-PGRN antibody raised against a synthetic peptide corresponding to the residues in the human PGRN protein at a dilution of 1:1,000 (EPR3781; Abcam, Cambridge, UK), or a mouse monoclonal anti-pS409/410 TDP-43 antibody raised against a phosphopeptide composed of CMDSKpSpSGWGM at a dilution of 1:500 (TIP-PTD-M01; Cosmo Bio, Tokyo, Japan).

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: The sections were then incubated at 4°C overnight with a rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 1 to 50 of the human TMEM106B protein at a concentration of 0.1 μg/ml (A303-439A; Bethyl Laboratories, Montgomery, TX, USA), a rabbit monoclonal anti-PGRN antibody raised against a synthetic peptide corresponding to the residues in the human PGRN protein at a dilution of 1:1,000 (EPR3781; Abcam, Cambridge, UK), or a mouse monoclonal anti-pS409/410 TDP-43 antibody raised against a phosphopeptide composed of CMDSKpSpSGWGM at a dilution of 1:500 (TIP-PTD-M01; Cosmo Bio, Tokyo, Japan).

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: The sections were then incubated at 4°C overnight with a rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 1 to 50 of the human TMEM106B protein at a concentration of 0.1 μg/ml (A303-439A; Bethyl Laboratories, Montgomery, TX, USA), a rabbit monoclonal anti-PGRN antibody raised against a synthetic peptide corresponding to the residues in the human PGRN protein at a dilution of 1:1,000 (EPR3781; Abcam, Cambridge, UK), or a mouse monoclonal anti-pS409/410 TDP-43 antibody raised against a phosphopeptide composed of CMDSKpSpSGWGM at a dilution of 1:500 (TIP-PTD-M01; Cosmo Bio, Tokyo, Japan).

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: The sections were then incubated at 4°C overnight with a rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 1 to 50 of the human TMEM106B protein at a concentration of 0.1 μg/ml (A303-439A; Bethyl Laboratories, Montgomery, TX, USA), a rabbit monoclonal anti-PGRN antibody raised against a synthetic peptide corresponding to the residues in the human PGRN protein at a dilution of 1:1,000 (EPR3781; Abcam, Cambridge, UK), or a mouse monoclonal anti-pS409/410 TDP-43 antibody raised against a phosphopeptide composed of CMDSKpSpSGWGM at a dilution of 1:500 (TIP-PTD-M01; Cosmo Bio, Tokyo, Japan).

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: The sections were then incubated at 4°C overnight with a rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 1 to 50 of the human TMEM106B protein at a concentration of 0.1 μg/ml (A303-439A; Bethyl Laboratories, Montgomery, TX, USA), a rabbit monoclonal anti-PGRN antibody raised against a synthetic peptide corresponding to the residues in the human PGRN protein at a dilution of 1:1,000 (EPR3781; Abcam, Cambridge, UK), or a mouse monoclonal anti-pS409/410 TDP-43 antibody raised against a phosphopeptide composed of CMDSKpSpSGWGM at a dilution of 1:500 (TIP-PTD-M01; Cosmo Bio, Tokyo, Japan).

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: The sections were then incubated at 4°C overnight with a rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 1 to 50 of the human TMEM106B protein at a concentration of 0.1 μg/ml (A303-439A; Bethyl Laboratories, Montgomery, TX, USA), a rabbit monoclonal anti-PGRN antibody raised against a synthetic peptide corresponding to the residues in the human PGRN protein at a dilution of 1:1,000 (EPR3781; Abcam, Cambridge, UK), or a mouse monoclonal anti-pS409/410 TDP-43 antibody raised against a phosphopeptide composed of CMDSKpSpSGWGM at a dilution of 1:500 (TIP-PTD-M01; Cosmo Bio, Tokyo, Japan).

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: The sections were then incubated at 4°C overnight with a rabbit polyclonal anti-TMEM106B antibody raised against a peptide spanning amino acid residues 1 to 50 of the human TMEM106B protein at a concentration of 0.1 μg/ml (A303-439A; Bethyl Laboratories, Montgomery, TX, USA), a rabbit monoclonal anti-PGRN antibody raised against a synthetic peptide corresponding to the residues in the human PGRN protein at a dilution of 1:1,000 (EPR3781; Abcam, Cambridge, UK), or a mouse monoclonal anti-pS409/410 TDP-43 antibody raised against a phosphopeptide composed of CMDSKpSpSGWGM at a dilution of 1:500 (TIP-PTD-M01; Cosmo Bio, Tokyo, Japan).

Techniques: Expressing, Immunohistochemistry, Staining