Journal: bioRxiv

Article Title: JAZF1-SUZ12 dysregulates PRC2 function and gene expression during cell differentiation

doi: 10.1101/2021.04.15.440052

Figure Lengend Snippet: a. Left: Immunoblots for FLAG, EZH2, FUS, β-actin and H3 in nucleoplasm (Np) and chromatin (Ch) fractions from mock or RNaseA-treated Suz12 GT/GT cells expressing either FLAG-tagged SUZ12, SUZ12Δ93 or JAZF1-SUZ12. Representative of three independent experiments. Right : Fold change in FLAG, EZH2, FUS and β-actin in the chromatin fraction upon RNaseA treatment (mean and s.d, n=3). b. As a., but for endogenous SUZ12, EZH2, JARID2, FUS, β-actin and H3 in Aebp2 WT/WT , Aebp2 GT/GT and Jarid2 GT/GT ESC. c. As a., but for endogenous SUZ12, EZH2, EPOP, FUS, β-actin and H3 in Epop WT/WT and Epop GT/GT ESC. See also .

Article Snippet: Proteins were detected with primary antibodies to FLAG M2 (Sigma, A8592), HA 3F10 (Roche, 12013819001), V5 (Abcam, ab15828), SUZ12 (Santa Cruz sc-46264), EZH2 (CST, 3147), JARID2 (CST, 13594), AEBP2 (CST 14129), PCL2 (Proteintech 16208-1-AP), EPOP (kind gift of L. Di Croce), TRRAP (Abcam, ab73546), FUS (Novus Biologicals 100-565), β-actin (CST 4967), alpha tubulin (CST 2144), H3K27me3 (Abcam ab192985) and H3 (Abcam ab1791) and HRP-conjugated secondary antibodies (anti-mouse (Dako, P0447) or anti-rabbit (Dako, P0448)).

Techniques: Western Blot, Expressing

Journal: bioRxiv

Article Title: JAZF1-SUZ12 dysregulates PRC2 function and gene expression during cell differentiation

doi: 10.1101/2021.04.15.440052

Figure Lengend Snippet: a. Left: Immunoblots for SUZ12, FUS, β-actin, and H3 in nucleoplasm (Np) and chromatin (Ch) fractions from mock or RNaseA-treated WT or Pali1 GT/GT cells. Representative of two independent experiments. Right : Fold change in FLAG, FUS and β-actin in the chromatin fraction upon RNaseA treatment (mean and s.d, n=2) b. Immunoblots for SUZ12, FUS, β-actin and H3 in nucleoplasm (Np) and chromatin (Ch) fractions from mock or RNaseA-treated Pcl2 GT/GT Pcl1/3 cKO cells pre-treated or not with 4-hydroxytamoxifen (tmx) to induce deletion of Pcl1 and Pcl3 (mean and s.d., n=2).

Article Snippet: Proteins were detected with primary antibodies to FLAG M2 (Sigma, A8592), HA 3F10 (Roche, 12013819001), V5 (Abcam, ab15828), SUZ12 (Santa Cruz sc-46264), EZH2 (CST, 3147), JARID2 (CST, 13594), AEBP2 (CST 14129), PCL2 (Proteintech 16208-1-AP), EPOP (kind gift of L. Di Croce), TRRAP (Abcam, ab73546), FUS (Novus Biologicals 100-565), β-actin (CST 4967), alpha tubulin (CST 2144), H3K27me3 (Abcam ab192985) and H3 (Abcam ab1791) and HRP-conjugated secondary antibodies (anti-mouse (Dako, P0447) or anti-rabbit (Dako, P0448)).

Techniques: Western Blot

Journal: Acta neuropathologica

Article Title: Transportin 1 accumulates specifically with FET proteins but no other transportin cargos in FTLD-FUS and is absent in FUS inclusions in ALS with FUS mutations.

doi: 10.1007/s00401-012-1020-6

Figure Lengend Snippet: Fig. 2 Co-localization of Trn1 and FUS in all FTLD-FUS subtypes. Double-label immunofluorescence for FUS (red) and Trn1 (green), with DAPI staining of nuclei in the merged images. FUS-positive inclusions in all FTLD-FUS subtypes consistently showed co-localization of FUS and Trn1, as shown for neuronal cytoplasmic inclusions (NCI) and neuronal intranuclear inclusions (NII, arrow in a) in the dentate granule cells in aFTLD-U (a), NCI in the temporal cortex of NIFID (b) and NCI and glial cytoplasmic inclusions in the spinal cord of BIBD (c). Scale bar 10 lm

Article Snippet: Table 3 Trn1 cargo proteins with PY-NLS investigated in FTLD-FUS Protein name Antibody Physiological staining pattern Inclusions in FTLD-FUS Company Dilution Type FUS ProteinTech Group (60160-1-Ig) 1:1,000 MM nucl pos Sigma (HPA008784) 1:2,000 RP nucl pos TAF15 Bethyl Laboratories (IHC-00094-1) 1:200 RP nucl pos EWS Santa Cruz (clone G5) 1:200 MM nucl [ cyto pos Bethyl Laboratories (IHC-00086) 1:200 RP nucl [ cyto pos hnRNP A1 Santa Cruz (clone 4B10) 1:500 MM nucl neg hnRNP A0 Abcam (ab66661) 1:100 RP nucl neg hnRNP A2/B1 Sigma-Aldrich (clone DP3B3) 1:500 MM nucl neg hnRNP M3/M4 Santa Cruz (clone 2A6) 1:250 MM nucl neg hnRNP D ProteinTech Group (12770-1-AP) 1:500 RP nucl neg hnRNP H1 ProteinTech Group (14774-1-AP) 1:50 RP nucl [ cyto neg PQBP-1 ProteinTech Group (16264-1-AP) 1:250 RP nucl neg SAM68 ProteinTech Group (10222-1-AP) 1:250 RP nucl neg SLM-2 ProteinTech Group (13563-1-AP) 1:50 RP nucl neg HEXIM1 ProteinTech Group (15676-1-AP) 1:50 RP nucl neg RBM39 ProteinTech Group (21339-1-AP) 1:50 RP nucl [ cyto neg HuR Santa Cruz (clone 3A2) 1:250 MM nucl neg PABPN1 Abcam (EP3000Y) 1:1,000 RM nucl neg Cyto cytoplasmatic, MM mouse monoclonal, nucl nuclear, pos positive, RP rabbit polyclonal, RM rabbit monoclonal, neg negative Trn1 staining in neurological controls The normal and neurological control cases did not reveal Trn1 immunoreactive pathology with one exception (Table 2).

Techniques: Staining

Journal: Acta neuropathologica

Article Title: Transportin 1 accumulates specifically with FET proteins but no other transportin cargos in FTLD-FUS and is absent in FUS inclusions in ALS with FUS mutations.

doi: 10.1007/s00401-012-1020-6

Figure Lengend Snippet: Fig. 3 Absence of Trn1 pathology in ALS-FUS. Double-label immuno- fluorescence for FUS (red) and Trn1 (green), with DAPI staining of nuclei in the merged images. FUS-positive inclusions in ALS-FUS were not labeled for Trn1 as shown for neuronal cytoplasmic inclusions in the spinal cord for three different FUS mutations (a–c). Note the physiological nuclear staining for Trn1 in inclusion bearing cells. FUS-positive glial cytoplasmic inclusions present in a subset of ALS-FUS cases also showed no co-labeling for Trn1 (arrow in a). Scale bar 10 lm

Article Snippet: Table 3 Trn1 cargo proteins with PY-NLS investigated in FTLD-FUS Protein name Antibody Physiological staining pattern Inclusions in FTLD-FUS Company Dilution Type FUS ProteinTech Group (60160-1-Ig) 1:1,000 MM nucl pos Sigma (HPA008784) 1:2,000 RP nucl pos TAF15 Bethyl Laboratories (IHC-00094-1) 1:200 RP nucl pos EWS Santa Cruz (clone G5) 1:200 MM nucl [ cyto pos Bethyl Laboratories (IHC-00086) 1:200 RP nucl [ cyto pos hnRNP A1 Santa Cruz (clone 4B10) 1:500 MM nucl neg hnRNP A0 Abcam (ab66661) 1:100 RP nucl neg hnRNP A2/B1 Sigma-Aldrich (clone DP3B3) 1:500 MM nucl neg hnRNP M3/M4 Santa Cruz (clone 2A6) 1:250 MM nucl neg hnRNP D ProteinTech Group (12770-1-AP) 1:500 RP nucl neg hnRNP H1 ProteinTech Group (14774-1-AP) 1:50 RP nucl [ cyto neg PQBP-1 ProteinTech Group (16264-1-AP) 1:250 RP nucl neg SAM68 ProteinTech Group (10222-1-AP) 1:250 RP nucl neg SLM-2 ProteinTech Group (13563-1-AP) 1:50 RP nucl neg HEXIM1 ProteinTech Group (15676-1-AP) 1:50 RP nucl neg RBM39 ProteinTech Group (21339-1-AP) 1:50 RP nucl [ cyto neg HuR Santa Cruz (clone 3A2) 1:250 MM nucl neg PABPN1 Abcam (EP3000Y) 1:1,000 RM nucl neg Cyto cytoplasmatic, MM mouse monoclonal, nucl nuclear, pos positive, RP rabbit polyclonal, RM rabbit monoclonal, neg negative Trn1 staining in neurological controls The normal and neurological control cases did not reveal Trn1 immunoreactive pathology with one exception (Table 2).

Techniques: Staining, Labeling

Journal: Acta neuropathologica

Article Title: Transportin 1 accumulates specifically with FET proteins but no other transportin cargos in FTLD-FUS and is absent in FUS inclusions in ALS with FUS mutations.

doi: 10.1007/s00401-012-1020-6

Figure Lengend Snippet: Fig. 5 Absence of selected other Trn1 cargos (hnRNP A1, SAM68 and PABPN1) in FTLD-FUS. Double-label immunofluorescence for FUS (red) and other Trn1 cargos with PY-NLS (hnRNP A1, SAM68 and PABPN1, respectively, green) with DAPI staining of nuclei in the merged images in FTLD-FUS. FUS-positive inclusions in FTLD-FUS as shown here in the dentate gyrus of aFTLD-U were not labeled for hnRNP A1 (a), SAM68 (b) and PABPN1 (c). Scale bar 10 lm

Article Snippet: Table 3 Trn1 cargo proteins with PY-NLS investigated in FTLD-FUS Protein name Antibody Physiological staining pattern Inclusions in FTLD-FUS Company Dilution Type FUS ProteinTech Group (60160-1-Ig) 1:1,000 MM nucl pos Sigma (HPA008784) 1:2,000 RP nucl pos TAF15 Bethyl Laboratories (IHC-00094-1) 1:200 RP nucl pos EWS Santa Cruz (clone G5) 1:200 MM nucl [ cyto pos Bethyl Laboratories (IHC-00086) 1:200 RP nucl [ cyto pos hnRNP A1 Santa Cruz (clone 4B10) 1:500 MM nucl neg hnRNP A0 Abcam (ab66661) 1:100 RP nucl neg hnRNP A2/B1 Sigma-Aldrich (clone DP3B3) 1:500 MM nucl neg hnRNP M3/M4 Santa Cruz (clone 2A6) 1:250 MM nucl neg hnRNP D ProteinTech Group (12770-1-AP) 1:500 RP nucl neg hnRNP H1 ProteinTech Group (14774-1-AP) 1:50 RP nucl [ cyto neg PQBP-1 ProteinTech Group (16264-1-AP) 1:250 RP nucl neg SAM68 ProteinTech Group (10222-1-AP) 1:250 RP nucl neg SLM-2 ProteinTech Group (13563-1-AP) 1:50 RP nucl neg HEXIM1 ProteinTech Group (15676-1-AP) 1:50 RP nucl neg RBM39 ProteinTech Group (21339-1-AP) 1:50 RP nucl [ cyto neg HuR Santa Cruz (clone 3A2) 1:250 MM nucl neg PABPN1 Abcam (EP3000Y) 1:1,000 RM nucl neg Cyto cytoplasmatic, MM mouse monoclonal, nucl nuclear, pos positive, RP rabbit polyclonal, RM rabbit monoclonal, neg negative Trn1 staining in neurological controls The normal and neurological control cases did not reveal Trn1 immunoreactive pathology with one exception (Table 2).

Techniques: Staining, Labeling

Journal: Molecular cell

Article Title: ULK1/2 Regulates Stress Granule Disassembly Through Phosphorylation and Activation of VCP/p97

doi: 10.1016/j.molcel.2019.03.027

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit anti-FUS , Bethyl , Cat# IHC-00074; RRID:AB_2247085.

Techniques: Virus, Recombinant, Western Blot, Software

Journal: Scientific Reports

Article Title: A directional 3D neurite outgrowth model for studying motor axon biology and disease

doi: 10.1038/s41598-021-81335-z

Figure Lengend Snippet: Antibodies and buffers used in this study.

Article Snippet: FUS , Novus Biologicals , NB100-2599 , 1:100 , Buffer 1.

Techniques: