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: Biochimica et biophysica acta. Molecular basis of disease

Article Title: TMEM106B knockdown exhibits a neuroprotective effect in Parkinson's disease models via regulating autophagy-lysosome pathway.

doi: 10.1016/j.bbadis.2024.167553

Figure Lengend Snippet: Fig. 1. TMEM106B expression are enhanced in the PD patients and PD models. TMEM106B mRNA (A) and protein (B) expression in the venous blood samples of 55 patients with PD was elevated. (C) TMEM106B levels in MPP+-treated SH-SY5Y cells were analyzed by western blot analysis (n = 10). (D) TMEM106B levels in brain SNpc of MPTP-treated and control mice were analyzed by western blot analysis. **P < 0.05; ***P < 0.001, vs. control group (n = 10). Data were analyzed using unpaired two-tailed Student’s t-test.

Article Snippet: The following antibodies were used for immunofluorescence: rabbit anti-TMEM106B antibody (20995–1-AP; Proteintech), rabbit anti-TFEB antibody (13372–1-AP; Proteintech), and Alexa 594- conjugated anti-rabbit IgG (111–585-144; Jackson ImmunoResearch Laboratories).

Techniques: Expressing, Western Blot, Control, Two Tailed Test

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: TMEM106B knockdown exhibits a neuroprotective effect in Parkinson's disease models via regulating autophagy-lysosome pathway.

doi: 10.1016/j.bbadis.2024.167553

Figure Lengend Snippet: Fig. 2. TMEM106B knockdown decreased the expression of α-syn and increased the expression of TDP43 in SH-SY5Y cells induced by MPP+. (A) The green fluo rescence of the selected cells infected with the LV-shNC and LV-shTMEM106B. (B) The TMEM106B protein levels in the SH-SY5Y cells from different treatment groups were detected by western blot analysis (n = 4). (C) The levels of TMEM106B and LAMP1 protein in the SH-SY5Y cells from different treatment groups were detected by immunofluorescence (TMEM106B: n = 6; LAMP1: n = 4). (D) The expression of TDP43 and α-syn in the SH-SY5Y cells from different treatment groups were detected by western blot (n = 4). Double standardization was applied to process data. *P < 0.05, **P < 0.005, ***P < 0.001, ****P < 0.0001, vs. control group; #P < 0.05, ##P < 0.005, ###P < 0.001, ####P < 0.0001, vs. LV-shTMEM106B + MPP+ group; ^P < 0.05, vs. LV-shNC group. Data were analyzed using two-way ANOVA, followed by Tukey’s post-hoc test.

Article Snippet: The following antibodies were used for immunofluorescence: rabbit anti-TMEM106B antibody (20995–1-AP; Proteintech), rabbit anti-TFEB antibody (13372–1-AP; Proteintech), and Alexa 594- conjugated anti-rabbit IgG (111–585-144; Jackson ImmunoResearch Laboratories).

Techniques: Knockdown, Expressing, Infection, Western Blot, Immunofluorescence, Control

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: TMEM106B knockdown exhibits a neuroprotective effect in Parkinson's disease models via regulating autophagy-lysosome pathway.

doi: 10.1016/j.bbadis.2024.167553

Figure Lengend Snippet: Fig. 3. Knockdown of TMEM106B activates the AMPK-mTOR-TFEB pathway in SH-SY5Y cells induced by MPP+. (A) The levels of TFEB in the cytosolic and nuclear fractions in the SH-SY5Y cells from different treatment groups were determined by western blot (n = 3). (B) Knockdown of TMEM106B promoted the translocation of endogenous TFEB to the nucleus which was determined by immunofluorescence (n = 6). (C) The levels of p-AMPK/AMPK, p-S6K1/S6K1, p-AKT/AKT and p-mTOR/ mTOR in the SH-SY5Y cells from different treatment groups were determined by western blot (p-AMPK/AMPK: n = 4; p-S6K1/S6K1: n = 3; p-AKT/AKT: n = 3; p- mTOR/mTOR: n = 6). Double standardization was applied to process data. *P < 0.05, **P < 0.005, ***P < 0.001, ****P < 0.0001, vs. control group; #P < 0.05, ##P < 0.005, ###P < 0.001, ####P < 0.0001, vs. LV-shTMEM106B + MPP+ group. Data were analyzed using two-way ANOVA, followed by Tukey’s post-hoc test.

Article Snippet: The following antibodies were used for immunofluorescence: rabbit anti-TMEM106B antibody (20995–1-AP; Proteintech), rabbit anti-TFEB antibody (13372–1-AP; Proteintech), and Alexa 594- conjugated anti-rabbit IgG (111–585-144; Jackson ImmunoResearch Laboratories).

Techniques: Knockdown, Western Blot, Translocation Assay, Immunofluorescence, Control

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: TMEM106B knockdown exhibits a neuroprotective effect in Parkinson's disease models via regulating autophagy-lysosome pathway.

doi: 10.1016/j.bbadis.2024.167553

Figure Lengend Snippet: Fig. 4. Knockdown of TMEM106B promoted ALP in SH-SY5Y cells induced by MPP+. (A) The levels of the LC3B-II, Beclin, and p62 in the SH-SY5Y cells from different treatment groups were determined by western blot. Representative blots are shown (P62, Beclin: n = 4; LC3B-II: n = 3). (B) The levels of the LAMP1, Pro- CTSD and mature-CTSD in the SH-SY5Y cells from different treatment groups were determined by western blot (LAMP1: n = 3; pro-CTSD, mature-CTSD: n = 4). (C) The lysosome contents in the SH-SY5Y cells from different treatment groups were determined by Lyso-Tracker Red staining (n = 8). Double standardization was applied to process data. *P < 0.05, ***P < 0.001, ****P < 0.0001, vs. control group; #P < 0.05, ##P < 0.005, ###P < 0.001, ####P < 0.0001, vs. LV-shTMEM106B + MPP+ group. Data were analyzed using two-way ANOVA, followed by Tukey’s post-hoc test.

Article Snippet: The following antibodies were used for immunofluorescence: rabbit anti-TMEM106B antibody (20995–1-AP; Proteintech), rabbit anti-TFEB antibody (13372–1-AP; Proteintech), and Alexa 594- conjugated anti-rabbit IgG (111–585-144; Jackson ImmunoResearch Laboratories).

Techniques: Knockdown, Western Blot, Staining, Control

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: TMEM106B knockdown exhibits a neuroprotective effect in Parkinson's disease models via regulating autophagy-lysosome pathway.

doi: 10.1016/j.bbadis.2024.167553

Figure Lengend Snippet: Fig. 5. Knockdown of TMEM106B attenuated apoptosis in SH-SY5Y cells induced by MPP+ (Bax: n = 5; Bcl-2: n = 4; Cleaved caspase-3: n = 3). Double stan dardization was applied to process data. *P < 0.05, **P < 0.005, ***P < 0.001, ****P < 0.0001, vs. control group; #P < 0.05, ##P < 0.005, vs. LV-shTMEM106B + MPP+ group. Data were analyzed using two-way ANOVA, followed by Tukey’s post-hoc test.

Article Snippet: The following antibodies were used for immunofluorescence: rabbit anti-TMEM106B antibody (20995–1-AP; Proteintech), rabbit anti-TFEB antibody (13372–1-AP; Proteintech), and Alexa 594- conjugated anti-rabbit IgG (111–585-144; Jackson ImmunoResearch Laboratories).

Techniques: Knockdown, Control

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: TMEM106B knockdown exhibits a neuroprotective effect in Parkinson's disease models via regulating autophagy-lysosome pathway.

doi: 10.1016/j.bbadis.2024.167553

Figure Lengend Snippet: Fig. 6. Knockdown of TMEM106B improved motor behavior and alleviated the loss of dopaminergic neuron in MPTP-treated mice. (A) Distribution of AAV-shNC and AAV-shTMEM106B (green-labeled) in TH positive neurons in the SN. (B) The TMEM106B protein levels in the SN from different treatment groups were detected by western blot analysis (n = 4). (C) In pole test, falling time was defined as the time required for the mouse to crawl back-and-forth on the rod (n = 15). (D) In rotarod test, latency to fall was defined as the time when mouse maintained the position on the rotating rod (n = 15). (E) Immunohistochemical staining of TH-positive neurons in the SN from different treatment groups (n = 6). (F) The TH protein levels in the SN from different treatment groups were detected by western blot analysis (n = 4). Double standardization was applied to process data. **P < 0.005, ***P < 0.001, ****P < 0.0001, vs. control group; #P < 0.05, ##P < 0.005, ###P < 0.001, ####P < 0.0001, vs. AAV-shTMEM106B + MPTP group. Data were analyzed using two-way ANOVA, followed by Tukey’s post-hoc test.

Article Snippet: The following antibodies were used for immunofluorescence: rabbit anti-TMEM106B antibody (20995–1-AP; Proteintech), rabbit anti-TFEB antibody (13372–1-AP; Proteintech), and Alexa 594- conjugated anti-rabbit IgG (111–585-144; Jackson ImmunoResearch Laboratories).

Techniques: Knockdown, Labeling, Western Blot, Immunohistochemical staining, Staining, Control

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: TMEM106B knockdown exhibits a neuroprotective effect in Parkinson's disease models via regulating autophagy-lysosome pathway.

doi: 10.1016/j.bbadis.2024.167553

Figure Lengend Snippet: Fig. 7. Knockdown of TMEM106B increased the expression of TDP43 and alleviated α-syn level and apoptosis in MPTP-induced PD mice. (A) The expression of TDP43 and α-syn in the SN from different treatment groups were detected by western blot (TDP43: n = 5; α-syn: n = 4). (B) The levels of the Bax, Bcl-2, and cleaved caspase-3 in the SN from different treatment groups were determined by western blot (Bax, Bcl-2: n = 4; Cleaved caspase-3: n = 3). Double standardization was applied to process data. *P < 0.05, **P < 0.005, ***P < 0.001, ****P < 0.0001, vs. control group; #P < 0.05, ##P < 0.005, ###P < 0.001, ####P < 0.0001, vs. AAV- shTMEM106B + MPTP group. Data were analyzed using two-way ANOVA, followed by Tukey’s post-hoc test.

Article Snippet: The following antibodies were used for immunofluorescence: rabbit anti-TMEM106B antibody (20995–1-AP; Proteintech), rabbit anti-TFEB antibody (13372–1-AP; Proteintech), and Alexa 594- conjugated anti-rabbit IgG (111–585-144; Jackson ImmunoResearch Laboratories).

Techniques: Knockdown, Expressing, Western Blot, Control

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: TMEM106B knockdown exhibits a neuroprotective effect in Parkinson's disease models via regulating autophagy-lysosome pathway.

doi: 10.1016/j.bbadis.2024.167553

Figure Lengend Snippet: Fig. 8. Knockdown of TMEM106B activated the AMPK-mTOR-TFEB pathway and enhanced ALP in MPTP-induced PD mice. (A) The levels of TFEB, p-AMPK/AMPK, p-S6K1/S6K1, p-AKT/AKT and p-mTOR/mTOR in the SN from different treatment groups were determined by western blot (TFEB n = 4; p-AMPK/AMPK: n = 4; p- S6K1/S6K1: n = 3; p-AKT/AKT: n = 3; p-mTOR/mTOR: n = 4). (B) The levels of the LC3B-II, p62, and Beclin in SN from different treatment groups were determined by western blot (P62, LC3B-II: n = 3; Beclin: n = 4). (C) The levels of the LAMP1, pro-CTSD and mature-CTSD in SN from different treatment groups were determined by western blot (LAMP1: n = 3; pro-CTSD, mature-CTSD: n = 5). Double standardization was applied to process data. *P < 0.05, **P < 0.005, ***P < 0.001, ****P < 0.0001, vs. control group; #P < 0.05, ##P < 0.005, ###P < 0.001, ####P < 0.0001, vs. AAV-shTMEM106B + MPTP group. Data were analyzed using two-way ANOVA, followed by Tukey’s post-hoc test.

Article Snippet: The following antibodies were used for immunofluorescence: rabbit anti-TMEM106B antibody (20995–1-AP; Proteintech), rabbit anti-TFEB antibody (13372–1-AP; Proteintech), and Alexa 594- conjugated anti-rabbit IgG (111–585-144; Jackson ImmunoResearch Laboratories).

Techniques: Knockdown, Western Blot, Control

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