Journal: eLife

Article Title: An open cortico-basal ganglia loop allows limbic control over motor output via the nigrothalamic pathway

doi: 10.7554/elife.49995

Figure Lengend Snippet: Figure 1. Trans-synaptic wild-type rabies tracing reveals both closed and open cortico-basal ganglia loops. (A) The strategy to label striatal neurons connecting to the cerebral cortex by Wt-RABV trans-synaptic retrograde tracing, and CTb-based non-trans-synaptic anterograde tracing for mapping cortico-striatal terminals. (B) Example image of Wt-RABV/CTb injection into M1 (left). After 66–70 hr of survival time, Wt-RABV was transfected up to 3rd- order neurons, which were found in various striatal subregions (right). Scale bars, 1 mm (left), 500 mm (right). (C) 3D-reconstruction of Wt-RABV+ striatal neurons from the M1 injection case shown in (B). The two different angles emphasize the presence of Wt-RABV+ neurons throughout all of the striatum (VS, DMS, DLS, and TS). (D) Schema of Wt-RABV/CTb injection in M1. (E) Anterogradely labeled CTb+ cortico-striatal terminals (green) and retrogradely labeled Wt-RABV+ striatal neurons (purple) from the M1 injection case shown in (B). (F) Density map showing the distribution of Wt-RABV+ neurons throughout the striatum from M1 injection. Black contours indicate approximate areas receiving cortico-striatal inputs from M1. Color maps indicate the intensity of Wt-RABV+ labeling. (G–I) The same analyses for Wt-RABV/CTb injection in M2. (J–L) The same analyses for Wt-RABV/CTb injection in mPFC. (M) Normalized distribution of Wt-RABV+ neurons across five striatal regions (VMS, VLS, DMS, DLS, TS) showing differences Figure 1 continued on next page

Article Snippet: DOI: https://doi.org/10.7554/eLife.49995 15 of 29 Continued Reagent type (species) or resource Designation Source or reference Identifiers Additional information Strain, strain background (recombinant rabies virus) EnvA.dGRabies.eGFP Salk Vector Core RRID: SCR_014847 Strain, strain background (adeno-associated virus) AAV9.FLEX.tdTomato University of Penn Viral Vector Core RRID: SCR_015406 Strain, strain background (adeno-associated virus) AAV9.FLEX.eGFP University of Penn Viral Vector Core RRID: SCR_015406 Strain, strain background (adeno-associated virus) AAV9.CAG.tdTomato UNC Viral Vector Core RRID: SCR_002448 Strain, strain background (adeno-associated virus) AAV9.hsyn.ChR2.eGFP University of Penn Viral Vector Core RRID: SCR_015406 Strain, strain background (adeno-associated virus) AAV5-EF1a-DIOhChR2(H134R)-mCherry University of Penn Viral Vector Core Antibody anti-wild type rabies phosphoprotein mouse monoclonal antibody commercially unavailable (Raux et al., 1997) (1:5000) Antibody anti-cholera toxin b-subunit goat polyclonal antibody List Biological Laboratories Cat.# 704 (1:15000) Antibody anti-tyrosine hydroxylase (TH) mouse monoclonal antibody Millipore Cat.# MAB318 (1:1000) Antibody anti-NeuN rabbit polyclonal antibody Abcam Cat.# ab104225 (1:1000) Antibody anti-GFP chicken polyclonal antibody Novus Biologicals Cat.# NB100-1614 (1:1000) Antibody anti-substance P mouse monoclonal antibody Abcam Cat.# ab14184 (1:1000) Antibody anti-mouse IgG horseradish peroxydase (HRP) (host: rabbit, polyclonal) DAKO Cat.# P260 (1:200) Antibody anti-goat IgG horseradish peroxydase (HRP) (host: rabbit, polyclonal) DAKO Cat.# P044901-2 (1:200) Antibody anti-mouse Alexa Fluor 488 (host: donkey, polyclonal) Jackson ImmunoResearch Laboratories Cat.# 715-545-151 (1:250) Antibody anti-mouse Cy3 (host: donkey, polyclonal) Jackson ImmunoReseach Laboratories Cat.# 715-165-151 (1:250) Antibody anti-mouse Cy5 (host: donkey, polyclonal) Jackson ImmunoReseach Laboratories Cat.# 715-175-151 (1:250) Continued on next page Aoki et al. eLife 2019;8:e49995.

Techniques: Retrograde Tracing, Anterograde Tracing, Injection, Transfection, Labeling

Journal: The American journal of pathology

Article Title: Increased expression of versican in the inflammatory response to UVB- and reactive oxygen species-induced skin tumorigenesis.

doi: 10.1016/j.ajpath.2011.08.042

Figure Lengend Snippet: Figure 3. Expression of continuously up-regulated genes after UVB expo- sure. A: Four genes were significantly up-regulated at both 3 and 24 hours after UVB exposure. Relative expression levels were determined by real-time quantitative PCR. Each of the two groups of RNAs isolated from wild-type and Ogg1 knockout mice was assayed in duplicate. Reactions were normal- ized to GAPDH expression levels. *P 0.05; **P 0.01; and ***P 0.001. B: Immunohistochemical study of versican expression after UVB irradiation. Versican is barely expressed in the wild-type mouse epidermis and dermis in the absence of UVB irradiation. At 24 hours after UVB exposure, versican was expressed in the wild-type epidermis. In Ogg1 knockout mice at 24 hours, versican was strongly expressed in the epidermis (arrowheads), as well as in dermal fibroblasts (arrows). Positive signals are seen as reddish-brown deposits produced on reaction with the 3-amino-9-ethylcarbazole substrate. Scale bar 30 m. C: Versican expression after UVB exposure, as deter- mined by Western blotting. Versican was more strongly up-regulated in Ogg1 knockout than in wild-type mice at 24 and 48 hours after UVB exposure. This up-regulation was time-dependent. The band at approximately 75 kDa indi- cates that the antibody for versican can detect the V1 isoform. /-Tubulin was used as the loading control. Data are representative of three separate determinations.

Article Snippet: Sections were incubated for 16 hours at 4°C with the following primary antibodies: rabbit polyclonal anti-mouse IL-1 (1:1000 dilution; Abcam, Cambridge, MA), rabbit polyclonal anti-mouse versican (1: 100 dilution; LifeSpan Biosciences, Seattle, WA), rabbit polyclonal anti-human versican (1:125 dilution; Atlas Antibodies, Stockholm, Sweden), or rabbit polyclonal antimouse p53 (CM5) (1:500 dilution; Leica Biosystems Newcastle, Newcastle upon Tyne, UK).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Isolation, Knock-Out, Immunohistochemical staining, Irradiation, Produced, Western Blot, Control

Journal: The American journal of pathology

Article Title: Increased expression of versican in the inflammatory response to UVB- and reactive oxygen species-induced skin tumorigenesis.

doi: 10.1016/j.ajpath.2011.08.042

Figure Lengend Snippet: Figure 4. Versican expression in developing skin tumors of chronically UVB-exposed mice. Representative histological sections of SCC tumors from wild-type and Ogg1 knockout mice red-colored staining positively for versi- can are shown, with a summary of versican-positive wild-type and Ogg1 knockout mouse tumors overall. *P 0.05 for the ratio of malignant tumor to total tumors analyzed for each genotype.

Article Snippet: Sections were incubated for 16 hours at 4°C with the following primary antibodies: rabbit polyclonal anti-mouse IL-1 (1:1000 dilution; Abcam, Cambridge, MA), rabbit polyclonal anti-mouse versican (1: 100 dilution; LifeSpan Biosciences, Seattle, WA), rabbit polyclonal anti-human versican (1:125 dilution; Atlas Antibodies, Stockholm, Sweden), or rabbit polyclonal antimouse p53 (CM5) (1:500 dilution; Leica Biosystems Newcastle, Newcastle upon Tyne, UK).

Techniques: Expressing, Knock-Out, Staining

Journal: The American journal of pathology

Article Title: Increased expression of versican in the inflammatory response to UVB- and reactive oxygen species-induced skin tumorigenesis.

doi: 10.1016/j.ajpath.2011.08.042

Figure Lengend Snippet: Figure 5. Versican expression in human skin tumors. A: Immunohistochemical study of versican expression in malignant (lentigo maligna melanoma, basal cell carcinoma, and squamous cell carcinoma) and benign (seborrheic keratosis and lentigo senilis) tumors. Arrowheads indicate the borders between the seborrheic keratosis tumor and the normal skin. Scale bar 100 m. B: Classification of versican expression in skin tumors, grouped as dermal and stromal versus tumoral.

Article Snippet: Sections were incubated for 16 hours at 4°C with the following primary antibodies: rabbit polyclonal anti-mouse IL-1 (1:1000 dilution; Abcam, Cambridge, MA), rabbit polyclonal anti-mouse versican (1: 100 dilution; LifeSpan Biosciences, Seattle, WA), rabbit polyclonal anti-human versican (1:125 dilution; Atlas Antibodies, Stockholm, Sweden), or rabbit polyclonal antimouse p53 (CM5) (1:500 dilution; Leica Biosystems Newcastle, Newcastle upon Tyne, UK).

Techniques: Expressing, Immunohistochemical staining

Journal: The American journal of pathology

Article Title: Increased expression of versican in the inflammatory response to UVB- and reactive oxygen species-induced skin tumorigenesis.

doi: 10.1016/j.ajpath.2011.08.042

Figure Lengend Snippet: Figure 7. Proposed relationships of versican in the inflammatory response leading to the development of skin tumors in terms of UVB-induced 8-oxoG accumulation. The accumulation of UVB/ROS-induced 8-oxoG in the skin leads to inflammatory reactions. High versican expression is induced by a highly inflammatory microenvironment with high numbers of infiltrated neu- trophils; conversely, neutrophil infiltration induces versican overexpression. More ROS will be generated at the inflammatory sites by neutrophils.

Article Snippet: Sections were incubated for 16 hours at 4°C with the following primary antibodies: rabbit polyclonal anti-mouse IL-1 (1:1000 dilution; Abcam, Cambridge, MA), rabbit polyclonal anti-mouse versican (1: 100 dilution; LifeSpan Biosciences, Seattle, WA), rabbit polyclonal anti-human versican (1:125 dilution; Atlas Antibodies, Stockholm, Sweden), or rabbit polyclonal antimouse p53 (CM5) (1:500 dilution; Leica Biosystems Newcastle, Newcastle upon Tyne, UK).

Techniques: Expressing, Over Expression, Generated

Journal: The American journal of pathology

Article Title: Increased expression of versican in the inflammatory response to UVB- and reactive oxygen species-induced skin tumorigenesis.

doi: 10.1016/j.ajpath.2011.08.042

Figure Lengend Snippet: Figure 6. Versican expression and neutrophil localization in human and murine skin tumors. A: Versican expression and inflammatory cells in human squamous cell carcinoma. Versican is strongly expressed in the dermal components (arrows) and inflammatory cells (arrowheads), seen at low magnification (left) and high magnification (right). Two focused areas are taken from different part of sections. Inflammatory cells, especially seg- mented leukocytes (neutrophils) (arrowheads), were strongly positive for versican. Scale bars 30 m. B: Versican expression and neutrophils in mouse skin. In the merged iamge, neutrophils with versican expression (arrows) appear yellow. Scale bar 30 m. C: Neutrophil infiltration after UVB irradiation in wild-type and Ogg1 knockout mice. Arrows indicate Gr-1-positive cells (green) in the dermis at 24 and 48 hours after UVB exposure in the wild-type and Ogg1 knockout mice. Scale bar 30 m. The accompanying graphs show the average number of neutrophils per 800 m2

Article Snippet: Sections were incubated for 16 hours at 4°C with the following primary antibodies: rabbit polyclonal anti-mouse IL-1 (1:1000 dilution; Abcam, Cambridge, MA), rabbit polyclonal anti-mouse versican (1: 100 dilution; LifeSpan Biosciences, Seattle, WA), rabbit polyclonal anti-human versican (1:125 dilution; Atlas Antibodies, Stockholm, Sweden), or rabbit polyclonal antimouse p53 (CM5) (1:500 dilution; Leica Biosystems Newcastle, Newcastle upon Tyne, UK).

Techniques: Expressing, Irradiation, Knock-Out

Journal: The Journal of biological chemistry

Article Title: Positive regulation of Raf1-MEK1/2-ERK1/2 signaling by protein serine/threonine phosphatase 2A holoenzymes.

doi: 10.1074/jbc.M502464200

Figure Lengend Snippet: FIGURE 2. RNAi-mediated knock-down of the PP2A regulatory subunit B. A, total RNA was isolated from multiple tetracycline-treated HEK T-Rex monoclonal cell lines stably expressing EV, B siRNA (B-KD), or epitope-tagged B subunit (B-FLAG or B-FLAG). Reverse transcription-PCR of the RNA was performed using primers specific for B, B, or GAPDH. Shown are ethidium bromide-stained 1.2% agarose gels of the result- ing PCR products. B, protein extracts (15 g) from the cell lines were analyzed by immunoblotting with B/ and GAPDH antibodies.

Article Snippet: IRDyeTM800- and AlexaFluor 680-conjugated fluorescent secondary antibodies were obtained from Rockland Immunochemicals (Gilbertsville, PA) andMolecularProbes (Eugene,OR), respectively.Commercial sources of other antibodies utilized in these studies were as follows: anti-phosphoRaf1(Ser-259), anti-phospho-MEK1/2(Ser-217 Ser-221), and anti-phospho-ERK1/2 (Thr-202 Tyr-204/Thr-185 Tyr-187) antibodies were from Cell Signaling Technology Inc. (Beverly, MA), anti-Raf1 (sc-133 and sc-7267) and anti-ERK1 (sc-93-G) antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was from Abcam, Inc. (Cambridge, MA), and anti-PP2A catalytic subunit antibody was from BD Biosciences Pharmingen.

Techniques: Knockdown, Isolation, Stable Transfection, Expressing, Reverse Transcription, Staining, Western Blot

Journal: The Journal of biological chemistry

Article Title: Positive regulation of Raf1-MEK1/2-ERK1/2 signaling by protein serine/threonine phosphatase 2A holoenzymes.

doi: 10.1074/jbc.M502464200

Figure Lengend Snippet: FIGURE 4. Activation of MEK1/2 and ERK1/2 in cells overexpressing PP2A B subunits. A, HEK T-Rex cells stably expressing EV, B-FLAG, or B-FLAG were treated with tetracycline for 24 h, serum-starved overnight, and then stimulated with EGF (50 ng/ml) for the indicated time points. Cell lysates (15 g of protein) were resolved by SDS-PAGE and immunoblotted with antibodies recognizing B/, p-ERK1/2, total ERK1/2, p-MEK1/2, and total MEK1/2. The data are repre- sentative of experiments that were performed at least three times. B, graphical representation of the data shown in A. The p-ERK1/2 and p-MEK1/2 signals were normalized to total ERK1/2 and MEK1/2 levels, respectively; the -fold activations are relative to normalized values of unstimulated cells expressing EV, which were set as 1.

Article Snippet: IRDyeTM800- and AlexaFluor 680-conjugated fluorescent secondary antibodies were obtained from Rockland Immunochemicals (Gilbertsville, PA) andMolecularProbes (Eugene,OR), respectively.Commercial sources of other antibodies utilized in these studies were as follows: anti-phosphoRaf1(Ser-259), anti-phospho-MEK1/2(Ser-217 Ser-221), and anti-phospho-ERK1/2 (Thr-202 Tyr-204/Thr-185 Tyr-187) antibodies were from Cell Signaling Technology Inc. (Beverly, MA), anti-Raf1 (sc-133 and sc-7267) and anti-ERK1 (sc-93-G) antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was from Abcam, Inc. (Cambridge, MA), and anti-PP2A catalytic subunit antibody was from BD Biosciences Pharmingen.

Techniques: Activation Assay, Stable Transfection, Expressing, SDS Page

Journal: The Journal of biological chemistry

Article Title: Positive regulation of Raf1-MEK1/2-ERK1/2 signaling by protein serine/threonine phosphatase 2A holoenzymes.

doi: 10.1074/jbc.M502464200

Figure Lengend Snippet: FIGURE 8. PP2A subunits co-immunoprecipitate with Raf1. Cells expressing EV, B-FLAG, B-FLAG, or B siRNA (B-KD) were unstimulated (A) or treated with or without EGF (B) prior to lysis in buffer B. A, FLAG immune complexes (FLAG IPs) were isolated from the clarified cell lysates and subjected to Western analysis using Raf1, FLAG, and PP2Ac antibodies. B, Raf1 immune complexes (Raf1 IPs) from lysates of cells treated with () or without () EGF were immunoblotted for Raf1, FLAG-B, and PP2Ac. An aliquot of each cell lysate was also probed for Raf1 and the indicated PP2A subunits using the corresponding antibodies (A and B, bottom panels).

Article Snippet: IRDyeTM800- and AlexaFluor 680-conjugated fluorescent secondary antibodies were obtained from Rockland Immunochemicals (Gilbertsville, PA) andMolecularProbes (Eugene,OR), respectively.Commercial sources of other antibodies utilized in these studies were as follows: anti-phosphoRaf1(Ser-259), anti-phospho-MEK1/2(Ser-217 Ser-221), and anti-phospho-ERK1/2 (Thr-202 Tyr-204/Thr-185 Tyr-187) antibodies were from Cell Signaling Technology Inc. (Beverly, MA), anti-Raf1 (sc-133 and sc-7267) and anti-ERK1 (sc-93-G) antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was from Abcam, Inc. (Cambridge, MA), and anti-PP2A catalytic subunit antibody was from BD Biosciences Pharmingen.

Techniques: Expressing, Lysis, Isolation, Western Blot

Journal: The Journal of biological chemistry

Article Title: Positive regulation of Raf1-MEK1/2-ERK1/2 signaling by protein serine/threonine phosphatase 2A holoenzymes.

doi: 10.1074/jbc.M502464200

Figure Lengend Snippet: FIGURE 9. PP2A-mediated dephosphorylation of Raf1. A, Raf1 immune complexes from cells treated with () or without () EGF were prepared in buffer A and then washed three times in phosphatase assay buffer containing or lacking 500 nM microcys- tin-LR (MC-LR). Dephosphorylation reactions were initiated by the addition of purified PP2AABCorABCholoenzymes(90ng)thathadbeenpretreatedwith()orwithout () 500 nM microcystin-LR. B and C, Raf1 or ERK1/2 immune complexes isolated from extracts of cells grown under normal conditions were incubated with ABC, ABC, or ABC holoenzymes (90 ng). FLAG eluate from EV-expressing cells was used as a con- trol. Following a 30-min incubation at 37 °C, the phosphatase reactions were terminated by the addition of SDS sample buffer and subjected to Western analysis using antibodies recognizing p-Raf1(Ser-259), total Raf1, p-ERK1/2, total ERK1/2, and PP2Ac.

Article Snippet: IRDyeTM800- and AlexaFluor 680-conjugated fluorescent secondary antibodies were obtained from Rockland Immunochemicals (Gilbertsville, PA) andMolecularProbes (Eugene,OR), respectively.Commercial sources of other antibodies utilized in these studies were as follows: anti-phosphoRaf1(Ser-259), anti-phospho-MEK1/2(Ser-217 Ser-221), and anti-phospho-ERK1/2 (Thr-202 Tyr-204/Thr-185 Tyr-187) antibodies were from Cell Signaling Technology Inc. (Beverly, MA), anti-Raf1 (sc-133 and sc-7267) and anti-ERK1 (sc-93-G) antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA), anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody was from Abcam, Inc. (Cambridge, MA), and anti-PP2A catalytic subunit antibody was from BD Biosciences Pharmingen.

Techniques: De-Phosphorylation Assay, Phosphatase Assay, Purification, Isolation, Incubation, Expressing, Western Blot

Journal: Annales De Pathologie

Article Title: Les lésions histologiques associées à l’infection par le SARS-CoV-2

doi: 10.1016/j.annpat.2020.12.009

Figure Lengend Snippet: Analyses immunohistochimiques anti-SARS. Cellules du parenchyme pulmonaire marquées avec, A : l’anticorps polyclonal de lapin anti-SARS Coronavirus (Invitrogen ; × 200), B : l’anticorps anti-SARS-CoV-2 (2019-nCoV) monoclonal de lapin ciblant la nucleoproteine virale (Sino Biological ; × 200), C : l’anticorps monoclonal recombinant ciblant la nucléocapside du SARS coronavirus (CorisBio, × 200), et D : l’anticorps anti-protéine spike du SARS-CoV-2 (clone 1A9, × 200).

Article Snippet: Cellules du parenchyme pulmonaire marquées avec, A : l’anticorps polyclonal de lapin anti-SARS Coronavirus (Invitrogen ; × 200), B : l’anticorps anti-SARS-CoV-2 (2019-nCoV) monoclonal de lapin ciblant la nucleoproteine virale (Sino Biological ; × 200), C : l’anticorps monoclonal recombinant ciblant la nucléocapside du SARS coronavirus (CorisBio, × 200), et D : l’anticorps anti-protéine spike du SARS-CoV-2 (clone 1A9, × 200).

Techniques: Recombinant

Journal: Nature Communications

Article Title: Acetylation-induced TDP-43 pathology is suppressed by an HSF1-dependent chaperone program

doi: 10.1038/s41467-017-00088-4

Figure Lengend Snippet: TDP-43 acetylation-mimic aggregates recruit the degradative machinery and promote mitochondrial dysfunction. a Cells expressing aggregate-prone TDP-43-ΔNLS-K145Q were analyzed by double-labeling using ubiquitin, 20S proteasome, or p62 antibodies, and counterstained with DAPI ( blue ). White arrows highlight co-localization of ubiquitin, 20S proteasome, or p62 with TDP-43 aggregates. b Mouse TA muscles electroporated with TDP-43-ΔNLS-K145Q for 14 days were similarly analyzed using ubiquitin, 20S proteasome, and p62 antibodies. White arrows highlight prominent co-localization of TDP-43 aggregates with ubiquitin, 20S proteasome, or p62 in mouse muscles. c , d Mouse tissues were analyzed by immunoblotting ( c ) or microscopy ( d ) using ubiquitin or p62 antibodies, as well as the mitochondrial panel of antibody markers (COXIV, TOM20 and Cytochrome c). Scale bar = 50 μm

Article Snippet: The following primary antibodies were used for IF analysis: rabbit polyclonal anti-GFP (FL, Santa Cruz Biotechnology), mouse monoclonal anti-GFP (B2, Santa Cruz Biotechnology), rabbit polyclonal anti-phospho(409/410)-TDP-43 (Proteintech), mouse mono, polyubiquitinylated conjugates (FK2, Enzo Life Sciences), rabbit polyclonal anti-HA (Y-11, Santa Cruz Biotechnology), rabbit polyclonal anti-proteasome 20S core subunit (Enzo Life Sciences), mouse monoclonal anti-SQSTM1(Abnova), mouse monoclonal anti-UBQLN2 (555–624, Abnova), rat monoclonal anti-HSF1(10H8, Enzo Life Sciences), rabbit polyclonal anti-Hsp40/Hdj1 (Enzo Life Sciences), goat polyclonal anti-Hsp27 (M-20, Santa Cruz Biotechnology), rat anti-mouse CD8 (eBioscience, 1:100).

Techniques: Expressing, Labeling, Western Blot, Microscopy

Journal: Nature Communications

Article Title: Acetylation-induced TDP-43 pathology is suppressed by an HSF1-dependent chaperone program

doi: 10.1038/s41467-017-00088-4

Figure Lengend Snippet: Chronic TDP-43 pathology that persists in skeletal muscles recapitulates aspects of sIBM pathology. Mouse TA muscles were mock treated (control) or electroporated with GFP-tagged TDP-43-WT or TDP-43-ΔNLS-K145Q for 22 days. a Muscle sections were analyzed by immunohistochemistry using a monoclonal TDP-43 antibody detecting the ectopically expressed TDP-43 construct (human-specific TDP-43 antibody) or a polyclonal TDP-43 antibody detecting total TDP-43 (endogenous mouse or ectopic human TDP-43). Both antibodies detected TDP-43 aggregates ( black asterisks and arrows ). Scale bar = 50 μm. Muscle sections were also analyzed by standard GFP fluorescence ( b ) or double-labeled with GFP ( green ) and CD8 ( red ) antibodies ( c ), while nuclei were stained with DAPI. Scale bar = 20 μm. d H&E and Gomori trichrome staining showed robust myofiber pathology associated with chronic TDP-43 aggregation ( white asterisks ). Scale bar = 50 μm

Article Snippet: The following primary antibodies were used for IF analysis: rabbit polyclonal anti-GFP (FL, Santa Cruz Biotechnology), mouse monoclonal anti-GFP (B2, Santa Cruz Biotechnology), rabbit polyclonal anti-phospho(409/410)-TDP-43 (Proteintech), mouse mono, polyubiquitinylated conjugates (FK2, Enzo Life Sciences), rabbit polyclonal anti-HA (Y-11, Santa Cruz Biotechnology), rabbit polyclonal anti-proteasome 20S core subunit (Enzo Life Sciences), mouse monoclonal anti-SQSTM1(Abnova), mouse monoclonal anti-UBQLN2 (555–624, Abnova), rat monoclonal anti-HSF1(10H8, Enzo Life Sciences), rabbit polyclonal anti-Hsp40/Hdj1 (Enzo Life Sciences), goat polyclonal anti-Hsp27 (M-20, Santa Cruz Biotechnology), rat anti-mouse CD8 (eBioscience, 1:100).

Techniques: Immunohistochemistry, Construct, Fluorescence, Labeling, Staining