Journal: Molecular and Clinical Oncology

Article Title: Effects of endoplasmic reticulum stress-mediated CREB3L1 on apoptosis of glioma cells

doi: 10.3892/mco.2022.2516

Figure Lengend Snippet: Expression of CREB3L1, GRP78 and CHOP in World Health Organization grade I-Ⅳ glioma tissues detected by immunohistochemistry (magnification, x400). Values are expressed as the mean ± standard deviation (n=3). **P<0.01 vs. grade I. CREB3L1, cyclic adenosine monophosphate responsive element binding protein 3 like 1; CHOP, C/EBP-homologous protein; GRP78, glucose-regulated protein, 78 kDa.

Article Snippet: Sections were then blocked in 5% goat serum (cat. no. C0265; Beyotime Institute of Biotechnology) in PBS at room temperature for 30 min, followed by incubation with primary antibody against CREB3L1 (rabbit; cat. no. 11235-2-AP; 1:100 dilution; ProteinTech Group), glucose-regulated protein, 78 kDa [GRP78, also known as heat shock protein family A (Hsp70) member 5] (rabbit; Bs-1219R; 1:200 dilution; BIOSS) and C/EBP-homologous protein (CHOP) (rabbit; Bs-20669R; 1:300 dilution; BIOSS) in working solution overnight at 4˚C.

Techniques: Expressing, Immunohistochemistry, Standard Deviation, Binding Assay

Journal: Molecular and Clinical Oncology

Article Title: Effects of endoplasmic reticulum stress-mediated CREB3L1 on apoptosis of glioma cells

doi: 10.3892/mco.2022.2516

Figure Lengend Snippet: Expression of CREB3L1, GRP78, CHOP and Bcl-2 following incubation with different concentrations of TG. Values are expressed as the mean ± standard deviation (n=3). *P<0.05, **P<0.01 vs. control. CREB3L1, cyclic adenosine monophosphate responsive element binding protein 3 like 1; CHOP, C/EBP-homologous protein; GRP78, glucose-regulated protein, 78 kDa; TG, thapsigargin.

Article Snippet: Sections were then blocked in 5% goat serum (cat. no. C0265; Beyotime Institute of Biotechnology) in PBS at room temperature for 30 min, followed by incubation with primary antibody against CREB3L1 (rabbit; cat. no. 11235-2-AP; 1:100 dilution; ProteinTech Group), glucose-regulated protein, 78 kDa [GRP78, also known as heat shock protein family A (Hsp70) member 5] (rabbit; Bs-1219R; 1:200 dilution; BIOSS) and C/EBP-homologous protein (CHOP) (rabbit; Bs-20669R; 1:300 dilution; BIOSS) in working solution overnight at 4˚C.

Techniques: Expressing, Incubation, Standard Deviation, Binding Assay

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: International Journal of Molecular Sciences

Article Title: Antipsychotic Drug Aripiprazole Protects Liver Cells from Oxidative Stress

doi: 10.3390/ijms23158292

Figure Lengend Snippet: Endoplasmic reticulum UPR. Logarithmic fold-change in mRNA expression of long-term treated Fao of ( a ) Bip ( n = 4) , ( b ) Bip (one-time antipsychotic treatment with H 2 O 2 for 24 h; n = 3), ( c ) Bip (tunicamycin control, TUN; n = 4), and ( f ) Ddit3 / Chop ( n = 3). Protein amounts in long-term treated Fao, standardized to the total protein acquired from Coomassie-stained gels of ( d ) BIP ( n = 3) and € BIP (tunicamycin control; n = 4) with corresponding blots. Data are presented as the mean ± standard deviation (SD) and for ( a , b , d , f ) analyzed with one-way ANOVA followed by Dunnett’s test, comparing all samples to the baseline control with no added H 2 O 2 (*); samples with 1.5 mM H 2 O 2 added are also compared to their untreated control at 1.5 mM H 2 O 2 ( + p ≤ 0.05, +++ p ≤ 0.001); samples with 3 mM H 2 O 2 added are compared to their untreated control at 3 mM H 2 O 2 ( ## p ≤ 0.01, ### p ≤ 0.001). ( c ) Kruskal–Wallis test analysis with Dunn’s multiple comparison test; ( e ) one-way ANOVA analysis with Bonferroni’s multiple comparison test comparing each tunicamycin control to its untreated control. ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001; n : number of biological replicates.

Article Snippet: The following TaqMan probes labeled with the FAM dye (Thermo Fisher Scientific) were used: the reference gene Rn18s (Rn03928990_g1), Nrf2/Nfe2l2 (Rn00582415_m1), Srxn1 (Rn04337926_g1), Sirt1 (Rn01428096_m1), Foxo3α (Rn01441087_m1), Bip/Hspa5 (Rn00565250_m1), Chop/Ddit3 (Rn00492098_g1), p53 (Rn00755717_m1), p21 (Rn00589996_m1), Gadd45α (Rn00577049_m1), Gadd45 β (Rn01452530_g1), Gadd45 γ (Rn01352550_g1), Icam1 (Rn00564227_m1), Hspd1 (Rn01441529_g1), Txn2 (Rn00584162_g1), Clpp (Rn01527475_m1), Ymel1l (Rn00586650_m1).

Techniques: Expressing, Control, Staining, Standard Deviation, Comparison

Journal: Cell Reports

Article Title: A Wars2 Mutant Mouse Model Displays OXPHOS Deficiencies and Activation of Tissue-Specific Stress Response Pathways

doi: 10.1016/j.celrep.2018.11.080

Figure Lengend Snippet: Heart-Specific Activation of the Integrated Stress Response Causes Increased Plasma FGF21 and Systemic Changes in Metabolism (A) Relative plasma FGF21 protein levels in male mice at 12 months of age. Wars2 V117L/V117L and Wars2 +/+ animal numbers were 5 and 9, respectively; mean ± SD. Unpaired t test. (B and C) Relative mRNA expression levels of (B) Fgf21 and (C) Atf4 in tissues from male mice at 12 months of age. Wars2 V117L/V117L and Wars2 +/+ animal numbers were 5 and 5, respectively; mean ± SD. Data were log transformed and analyzed using t tests or a Mann-Whitney test ( Fgf21 in heart and Atf4 in skeletal muscle and iWAT). Fgf21 RNA expression is very low in kidney and in wild-type skeletal muscle: mean C T > 33. (D and E) Immunoblot analysis (D) and quantification (E) of p-eIF2α and total EIF2α protein levels in heart, liver, kidney, and skeletal muscle from female mice at 12 months of age. Wars2 V117L/V117L and Wars2 +/+ animal numbers were 3 and 3, respectively; bands were normalized to tubulin and expressed relative to wild-type as the mean ± SD. Significance was determined using an unpaired t test with Welch’s correction. (F and G) Relative mRNA expression levels of (F) Atf5 and (G) Chop in male Wars2 V117L/V117L and Wars2 +/+ tissues harvested at 12 months of age. Wars2 V117L/V117L and Wars2 +/+ animal numbers were 8 and 8, respectively; mean ± SD. Data were log transformed and analyzed using a t test or a Mann-Whitney test ( Atf5 and Chop in kidney and skeletal muscle). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Wars2 V117L/V117L mice are blue squares, and Wars2 +/+ mice are black triangles. See also and .

Article Snippet: Taqman probes used in this study: Wars2 (Exon 2-3) ( Mm04208965_m1), Wars2 (Exon 4-5) ( Mm04208967_m1), Wars2 (Exon 5-6) ( Mm00840490_m1), Pgc1α ( Mm01208835_m1), Atf4 ( Mm00515324_m1), Atf5 ( Mm00459515_m1), Chop ( Mm01135937_g1), Fgf21 ( Mm00840165_g1), Tfam ( Mm00447485_m1), Pparα ( Mm00440939_m1), Ucp1 (Mm01244861_m1), Dio2 ( Mm00515664_m1), Cidea ( Mm00432554_m1), Pparγ ( Mm00440945_m1), Cox7a1 ( Mm00438297_g1) and Cox8b ( Mm00432648_m1).

Techniques: Activation Assay, Clinical Proteomics, Expressing, Transformation Assay, MANN-WHITNEY, RNA Expression, Western Blot