Journal: Stem cell research & therapy

Article Title: Mature human induced pluripotent stem cell-derived cardiomyocytes promote angiogenesis through alpha-B crystallin.

doi: 10.1186/s13287-023-03468-4

Figure Lengend Snippet: Fig. 6 CRYAB was identified as an angiogenic factor of D56-CMs. a Scatter plot of RNA-seq data of D28-CMs and D56-CMs. The red line represents fourfold upregulation in D56-CMs compared with D28-CMs. b qRT-PCR analysis validated the upregulation of CRYAB in D56-CMs. The value for an adult heart sample was set to 1 as a reference. n = 3 per group. c Western blot analysis also confirmed the upregulation of CRYAB protein in D56-CMs compared with D28-CMs. GAPDH protein was used as a loading control. n = 3 per group. d Western blot analysis confirmed the KD of CRYAB (CRYAB-KD) after siRNA treatment in D56-CMs. As a control, non-targeting siRNA was used. n = 3 per group. e CRYAB-KD inhibited HUVEC migration by D56-CMs. The ratio of migrated cells in three independent experiments (n = 3–6 per experiment) is shown. f CRYAB-KD inhibited HUVEC tube lengths by D56-CMs. n = 4 per group. All data are the mean ± SEM, and the P values were determined with unpaired t tests (*P < 0.05, **P < 0.01)

Article Snippet: The Human CRYAB ELISA kit (CUSABIO, Houston, TX, USA; CSB-EL006008HU) was used according to the manufacturer’s protocol.

Techniques: RNA Sequencing, Quantitative RT-PCR, Western Blot, Control, Migration

Journal: Stem cell research & therapy

Article Title: Mature human induced pluripotent stem cell-derived cardiomyocytes promote angiogenesis through alpha-B crystallin.

doi: 10.1186/s13287-023-03468-4

Figure Lengend Snippet: Fig. 7 CRYAB-overexpressing D28-CMs enhanced angiogenesis in vivo. a qRT-PCR analysis validated AAV-mediated CRYAB overexpression (CRYAB-OE) 5 days after infection. An AAV vector carrying only tdTomato (tdTomato-OE) was used as a control. n = 4 per group. The value for an adult heart sample was set to 1 as a reference. b Immunostaining also confirmed significant upregulation of CRYAB in CRYAB-OE grafts (left) compared to tdTomato-OE grafts (right). Grafts are indicated by dotted lines. Scale bars, 100 µm. c Representative images of CD31+ microvessel (green) formation in βMHC+ grafts (red) at 4 weeks post-transplantation. Scale bars, 50 μm. d Quantification of microvessel formation in AAV-infected D28-CM grafts. Five sites were randomly selected from each animal. n = 4 per group. All data are the mean ± SEM, and the P values were determined with unpaired t tests (*P < 0.01, ***P < 0.001)

Article Snippet: The Human CRYAB ELISA kit (CUSABIO, Houston, TX, USA; CSB-EL006008HU) was used according to the manufacturer’s protocol.

Techniques: In Vivo, Quantitative RT-PCR, Over Expression, Infection, Plasmid Preparation, Control, Immunostaining, Transplantation Assay

Journal: Angiogenesis

Article Title: αB-crystallin/HspB5 regulates endothelial–leukocyte interactions by enhancing NF-κB-induced up-regulation of adhesion molecules ICAM-1, VCAM-1 and E-selectin

doi: 10.1007/s10456-013-9367-4

Figure Lengend Snippet: Ectopic expression of αB-crystallin increases E-selectin levels in HUVEC. a Representative Western Blot for αB-crystallin in HUVEC transduced with a lentiviral vector coding for full-length human αB-crystallin ( pgk:cryab ) or an empty control vector ( pgk:ev ). Actin served as loading control. b Representative FACS plots of E-selectin staining on HUVEC transduced with pgk:cryab and pgk:ev after treatment with TNF-α for 24 h. c Quantification of surface expression of E-selectin in HUVEC transduced with pgk:cryab ( white bars ) or empty vector controls ( black bars ). Cells were treated with TNF-α for 5 h ( left panel ) or 24 h ( right panel ) d HUVEC transduced with pgk:cryab ( white bars ) or empty vector controls ( black bars ) were stimulated for 3, 6, 12 and 24 h with TNF-α and expression of E-selectin relative to hprt was determined by qPCR ( Bars represent mean ± SD fold expression compared to pgk:ev after 3 h of TNF-α (normalized data from 3 independent experiments), * = p < 0.05). e HUVEC transduced with pgk:cryab ( white bars ) or empty vector controls ( black bars ) were grown to confluency and stimulated with TNF-α for 24 h before co-incubation with Jurkat cells for 15 min. Firmly adherent cells were microscopically quantified. ( Bars represent mean ± SD (normalized data from 3 individual independent), * = p < 0.05)

Article Snippet: For synthesis of the lentivirus vector pgk:cryab , a full length clone of human cryab cDNA (NM_001885.1, Origene, Rockville, MD) was cloned into a modified lentiviral pgk vector [ ] containing an internal ribosomal entry site (IRES) and eGFP as selection marker.

Techniques: Expressing, Western Blot, Transduction, Plasmid Preparation, Control, Staining, Incubation

Journal: Angiogenesis

Article Title: αB-crystallin/HspB5 regulates endothelial–leukocyte interactions by enhancing NF-κB-induced up-regulation of adhesion molecules ICAM-1, VCAM-1 and E-selectin

doi: 10.1007/s10456-013-9367-4

Figure Lengend Snippet: TNF-α-induced endothelial activation is reduced in endothelial cells derived from cryab -/- mice. a Representative western blot showing αB-crystallin expression in MyEnd wild type and MyEnd cryab - / - cells. b , c , d MyEnd wild type and MyEnd cryab - / - cells were treated with TNF-α for 3, 6, 18 and 24 h and expression of E-selectin ( b ), ICAM-1 ( c ) and VCAM-1 ( d ) relative to hprt was determined by qPCR. ( Bars represent mean ± SD (normalized data from 3 independent experiments), * = p < 0.05)

Article Snippet: For synthesis of the lentivirus vector pgk:cryab , a full length clone of human cryab cDNA (NM_001885.1, Origene, Rockville, MD) was cloned into a modified lentiviral pgk vector [ ] containing an internal ribosomal entry site (IRES) and eGFP as selection marker.

Techniques: Activation Assay, Derivative Assay, Western Blot, Expressing

Journal: Angiogenesis

Article Title: αB-crystallin/HspB5 regulates endothelial–leukocyte interactions by enhancing NF-κB-induced up-regulation of adhesion molecules ICAM-1, VCAM-1 and E-selectin

doi: 10.1007/s10456-013-9367-4

Figure Lengend Snippet: TNF-α-induced activation of NF-κB is reduced in the absence of αB-crystallin. MyEnd wt and MyEnd cryab - / - cells were treated with TNF-α for 15, 30, 45 min and 1 h, protein lysates were prepared and expression of IκB was determined by western blot. One representative blot of three independent experiments is shown

Article Snippet: For synthesis of the lentivirus vector pgk:cryab , a full length clone of human cryab cDNA (NM_001885.1, Origene, Rockville, MD) was cloned into a modified lentiviral pgk vector [ ] containing an internal ribosomal entry site (IRES) and eGFP as selection marker.

Techniques: Activation Assay, Expressing, Western Blot

Journal: Angiogenesis

Article Title: αB-crystallin/HspB5 regulates endothelial–leukocyte interactions by enhancing NF-κB-induced up-regulation of adhesion molecules ICAM-1, VCAM-1 and E-selectin

doi: 10.1007/s10456-013-9367-4

Figure Lengend Snippet: In vivo leukocyte–endothelial interactions are altered in cryab - / - mice, associated with decreased expression of endothelial adhesion molecules. Intravital microscopy was used to analyze leukocyte–endothelial interactions in inflamed venules of the cremaster muscle of wild type ( white bars ) and cryab - / - mice ( black bars ). Analysis of rolling velocity ( a ) and rolling flux ( b ) 3.5, 4, and 4.5 h after intrascrotal injection of TNF-α (n = 5, mean ± SD, * p < 0.05). qPCR analysis of mRNA expression of ICAM-1 ( c ), VCAM-1 ( d ) and E-selectin ( e ) relative to hprt in mouse cremaster muscles harvested 4.5 h after intrascrotal injection of TNF-α (n = 4, mean ± SD, * p < 0.05)

Article Snippet: For synthesis of the lentivirus vector pgk:cryab , a full length clone of human cryab cDNA (NM_001885.1, Origene, Rockville, MD) was cloned into a modified lentiviral pgk vector [ ] containing an internal ribosomal entry site (IRES) and eGFP as selection marker.

Techniques: In Vivo, Expressing, Intravital Microscopy, Injection, Muscles

Journal: Scientific reports

Article Title: Gestational high-sucrose diet mediated vascular hyper-contractility in mesenteric arteries from offspring.

doi: 10.1038/s41598-025-93361-2

Figure Lengend Snippet: Fig. 4. The RNA-Seq analysis of MAs from offspring. A Heat map of RNA-Seq. CON: n = 3, HS: n = 3. B Volcano map of RNA-Seq. CON: n = 3, HS: n = 3. C The mRNA expressions of APCDD1, HSPE1 and CRYAB. D The protein expressions of APCDD1, HSPE1 and CRYAB. E KEGG pathway enrichment analyses. n = 3–7 from different litters. *p < 0.05, **p < 0.001, ****p < 0.0001.

Article Snippet: Following blocking non-specific binding sites by 5% skim milk, the PVDF membranes were incubated with the primary antibodies, including GAPDH (1:2000, Affinity, China), β-actin (1:5000, Affinity, China), Wnt5a (1:1000, Affinity, China), PKCα (1:1000, Abcam, US), CRYAB (1:1000, Absin, China), HSPE1 (1:1000, Absin, China), and APCDD1 (1:1000, Proteintech, China).

Techniques: RNA Sequencing

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: αB-Crystallin Is a Target for Adaptive Immune Responses and a Trigger of Innate Responses in Preactive Multiple Sclerosis Lesions

doi: 10.1097/nen.0b013e3181e4939c

Figure Lengend Snippet: FIGURE 1. Serum immunoglobulin G (IgG) reactivity against total myelin protein extracts. Serum IgG reactivities against myelin proteins in 9 multiple sclerosis (MS) patients and 9 healthy controls were probed by Western blotting. In all cases, the a lanes contained 100-Kg samples of total myelin proteins, as characterized by Coomassie staining on the left. The b lanes contained 100-Kg samples that were supplemented with an extra 2 Kg of both recombinant human >B-crystallin (CRYAB) and the recombinant extracellular domain 1-125 of human myelin-oligodendrocyte glycoprotein (MOG). The increased reactivities (denser bands) at the 22-kd position in the b lanes indicate that CRYAB is the target of the antibody responses seen in the a lanes. MBP, myelin basic protein.

Article Snippet: Cytokine and chemokine profiling of culture supernatants was performed 24 hours after stimulation with 50 Kg/mL recombinant human CRYAB using a commercially available cytokine/chemokine antibody array (RayBiotech, Inc, Norcross, GA) according to the manufacturer’s instructions.

Techniques: Western Blot, Staining, Recombinant

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: αB-Crystallin Is a Target for Adaptive Immune Responses and a Trigger of Innate Responses in Preactive Multiple Sclerosis Lesions

doi: 10.1097/nen.0b013e3181e4939c

Figure Lengend Snippet: FIGURE 2. Multiple sclerosis (MS)Yinduced accumulation of >B- crystallin (CRYAB) in myelin preparations. Samples of 100 Kg of proteins from control myelin (CON) or myelin purified from tissue samples containing active MS lesions (MS) were probed by Western blotting using a mixture of a CRYAB-specific and 2¶,3¶-cyclic nucleotide 3¶-phosphodiesterase (CNPase)Yspecific monoclonal antibodies to verify equivalent loading of gels. Antibody-binding signals were compared with those generated against reference amounts of recombinant human CRYAB included on the same blot to permit absolute quantification (graph). CRYAB represents about 0.2% of all proteins in control myelin, but in MS-affected tissue samples, this level is approx- imately 10-fold higher. Figure 2 is representative of data obtained from 3 different MS samples.

Article Snippet: Cytokine and chemokine profiling of culture supernatants was performed 24 hours after stimulation with 50 Kg/mL recombinant human CRYAB using a commercially available cytokine/chemokine antibody array (RayBiotech, Inc, Norcross, GA) according to the manufacturer’s instructions.

Techniques: Control, Western Blot, Bioprocessing, Binding Assay, Generated, Recombinant

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: αB-Crystallin Is a Target for Adaptive Immune Responses and a Trigger of Innate Responses in Preactive Multiple Sclerosis Lesions

doi: 10.1097/nen.0b013e3181e4939c

Figure Lengend Snippet: FIGURE 4. >B-crystallin (CRYAB) in preactive lesions. (AYD) Close association of CRYAB and HLA-DR immunoreactivity in 2 exam- ples of preactive lesions (AYC and D). Original magnification: (AYD) 20.

Article Snippet: Cytokine and chemokine profiling of culture supernatants was performed 24 hours after stimulation with 50 Kg/mL recombinant human CRYAB using a commercially available cytokine/chemokine antibody array (RayBiotech, Inc, Norcross, GA) according to the manufacturer’s instructions.

Techniques:

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: αB-Crystallin Is a Target for Adaptive Immune Responses and a Trigger of Innate Responses in Preactive Multiple Sclerosis Lesions

doi: 10.1097/nen.0b013e3181e4939c

Figure Lengend Snippet: FIGURE 5. >B-crystallin (CRYAB) exclusively accumulates in oligodendrocytes in preactive multiple sclerosis (MS) lesions. (A, B) CRYAB is expressed in olig2-positive oligodendrocytes (arrows [A]) surrounded by intact myelin indicated by staining for myelin proteolipid protein ([PLP] B). (C, D) There is no colocalization of CRYAB with the astrocyte marker glial fibrillary acidic protein ([GFAP] C) or with the axonal marker neurofilament light ([NF-L] D). Original magnification: (AYD) 20.

Article Snippet: Cytokine and chemokine profiling of culture supernatants was performed 24 hours after stimulation with 50 Kg/mL recombinant human CRYAB using a commercially available cytokine/chemokine antibody array (RayBiotech, Inc, Norcross, GA) according to the manufacturer’s instructions.

Techniques: Staining, Marker

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: αB-Crystallin Is a Target for Adaptive Immune Responses and a Trigger of Innate Responses in Preactive Multiple Sclerosis Lesions

doi: 10.1097/nen.0b013e3181e4939c

Figure Lengend Snippet: FIGURE 6. >B-crystallin (CRYAB) is in contact with activated microglia in preactive lesions. (A, B) Colocalization of accumulated CRYAB and HLA-DR on microglia in crescent-shaped areas, suggesting direct contact between microglia and intact oligoden- drocytes. (C) On other occasions, there is colocalization in small intracellular vesicles, suggesting CRYAB uptake in the HLA- DRYcontaining endosomal compartment of microglia. Original magnification: (AYC) 20.

Article Snippet: Cytokine and chemokine profiling of culture supernatants was performed 24 hours after stimulation with 50 Kg/mL recombinant human CRYAB using a commercially available cytokine/chemokine antibody array (RayBiotech, Inc, Norcross, GA) according to the manufacturer’s instructions.

Techniques:

Journal: Journal of Neuropathology & Experimental Neurology

Article Title: αB-Crystallin Is a Target for Adaptive Immune Responses and a Trigger of Innate Responses in Preactive Multiple Sclerosis Lesions

doi: 10.1097/nen.0b013e3181e4939c

Figure Lengend Snippet: FIGURE 8. >B-crystallin (CRYAB) does not directly activate any toll-like receptor (TLR) known to be involved in recognizing bacterial products. To verify that activation of microglia by recombinant human CRYAB was not caused by bacterial contaminants in the preparation, CRYAB was tested for its ability to activate a panel or HEK293 reporter cells, each expressing a different TLR family member. In all cases, CRYAB up to 100 Kg/mL did not trigger any significant response. Results are the mean T SD of 3 experiments.

Article Snippet: Cytokine and chemokine profiling of culture supernatants was performed 24 hours after stimulation with 50 Kg/mL recombinant human CRYAB using a commercially available cytokine/chemokine antibody array (RayBiotech, Inc, Norcross, GA) according to the manufacturer’s instructions.

Techniques: Activation Assay, Recombinant, Expressing

Journal: Communications Biology

Article Title: CRYAB_K92 lactylation drives hypertrophy of the ligamentum flavum via an S100A16/RAGE-mediated glycolysis-fibrosis positive feedback loop

doi: 10.1038/s42003-026-10103-4

Figure Lengend Snippet: A The quantitative results of the Pearson’s Correlation Coefficient (PCC). B The Violin plot showing the distribution and differences in the intensity values of modification sites among different samples. C The expression heatmap of differentially modified sites, where red represents high expression, blue represents low expression, and gray represents non-quantifiable values in the corresponding sample. D–G The GO and KEGG pathway enrichment analyses of lactylation-activated proteins are presented via bubble plots (where D: MF, molecular function; E: BP, biological process; F: CC, cellular component; G: pathway enrichment). H Functional annotation of identified proteins. I The Volcano plot of differentially modified proteins and sites between LF/HLF groups. The information of the top five differentially modified sites is also marked in the figure. J The MS/MS spectra including C-terminal y-ions and N-terminal b-ions of CRYAB_K92.

Article Snippet: Recombinant S100A16 (MedChemExpress) was immobilized on a CM5 sensor chip, and serially diluted recombinant CRYAB (MedChemExpress) was injected as the analyte.

Techniques: Modification, Expressing, Functional Assay, Tandem Mass Spectroscopy

Journal: Communications Biology

Article Title: CRYAB_K92 lactylation drives hypertrophy of the ligamentum flavum via an S100A16/RAGE-mediated glycolysis-fibrosis positive feedback loop

doi: 10.1038/s42003-026-10103-4

Figure Lengend Snippet: A The co-IP showed the lactylation level of CRYAB in LF/HLF tissue. B The co-IP showed the regulation of exogenous lactate on the lactylation level of CRYAB in HLF cells. C The co-IP showed the lactylation level of CRYAB in HLF cells before and after the mutation at the CRYAB _K92 site. D , E The western blot and the quantification analysis showed the regulatory effect of exogenous lactate combined with the mutation at the CRYAB _ Kla92 on the fibrosis level in HLF cells. F , G The western blot and the quantification analysis showed the efficiency of P300 silencing and overexpression. H The co-IP showed the regulation of cellular lactylation level of CRYAB by the site mutation in combination with the lactylation-modifying enzyme P300 and the P300 inhibitor C646. I , J The western blot and the quantification analysis showed the regulatory effects on the fibrosis level of HLF cells before and after CRYAB _K92 mutation, as well as in combination with P300 and C646. K–M The results of EDU and FCM assays showing the regulation of the proliferation and cell cycle of HLF cells by the mutation at the CRYAB _K92 site in combination with P300 and C646. Scale bar=50 μm. O The co-IP results compared the regulatory effects of K92 site mutation, P300 knockout, and the use of the P300 inhibitor C646 on CRYAB lactylation in HLF cells. P , Q The western blot and the quantification analysis results compared the regulatory effects of K92 site mutation, P300 knockout, and the use of the P300 inhibitor C646 on the fibrosis level of HLF cells. The quantitative data are presented as mean ± SD, with individual data points overlaid to show the distribution. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Recombinant S100A16 (MedChemExpress) was immobilized on a CM5 sensor chip, and serially diluted recombinant CRYAB (MedChemExpress) was injected as the analyte.

Techniques: Co-Immunoprecipitation Assay, Mutagenesis, Western Blot, Over Expression, Knock-Out

Journal: Communications Biology

Article Title: CRYAB_K92 lactylation drives hypertrophy of the ligamentum flavum via an S100A16/RAGE-mediated glycolysis-fibrosis positive feedback loop

doi: 10.1038/s42003-026-10103-4

Figure Lengend Snippet: A The scatter plot shows differentially expressed proteins following mutation of the Kla92 site. S100A16 expression was significantly downregulated after the mutation. B Molecular docking prediction between S100A16 and CRYAB. The binding pose with the lowest PIPER pose energyis displayed. C The sensorgram of Surface plasmon resonance (SPR) analysis confirms a specific and direct interaction between S100A16 and CRYAB. D , E The western blot and quantitative analysis show that the protein half-life of S100A16 was significantly shortened after the CRYAB_K92 mutation. F The qRT-PCR results showed the mutation of the CRYAB _K92 site significantly reduced the mRNA expression level of S100A16 in HLF cells. G Actinomycin D chase analysis of S100A16 mRNA stability in CRYAB _K92 WT and Mut cells. mRNA levels at each time point were normalized to the respective 0-hour value (set as 100%). H , I The cellular immunofluorescence showed that after the CRYAB _K92 mutation, the signal levels of both lactylation (Kla, red) and S100A16 (green) decreased in HLF cells. The merged image shows co-localization between Kla and S100A16 signals. Nuclei were counterstained with DAPI (blue). Scale bar = 50 μm. J , K The cellular immunofluorescence showed that the mutation of CRYAB _K92 led to reduced expression levels of both CRYAB (red) and S100A16 (green) in HLF cells. Nuclei were counterstained with DAPI (blue). Scale bar = 50 μm. L The tissue immunofluorescence analysis showed that compared with LF tissue, S100A16 (red) expression was significantly increased in HLF tissue. Nuclei were counterstained with DAPI (blue). Scale bar = 50 μm. M The co-IP assays using a CRYAB antibody confirmed that the CRYAB _K92 mutation significantly reduced the level of S100A16 protein that interacts with CRYAB. The quantitative data are presented as mean ± SD, with individual data points overlaid to show the distribution. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Recombinant S100A16 (MedChemExpress) was immobilized on a CM5 sensor chip, and serially diluted recombinant CRYAB (MedChemExpress) was injected as the analyte.

Techniques: Mutagenesis, Expressing, Binding Assay, SPR Assay, Western Blot, Quantitative RT-PCR, Immunofluorescence, Co-Immunoprecipitation Assay

Journal: Communications Biology

Article Title: CRYAB_K92 lactylation drives hypertrophy of the ligamentum flavum via an S100A16/RAGE-mediated glycolysis-fibrosis positive feedback loop

doi: 10.1038/s42003-026-10103-4

Figure Lengend Snippet: A , B The results of western blot analysis showed the regulation of CRYAB_K92 in cooperation with S100A16 on the fibrosis and the RAGE pathway. C–F The results of EDU and FCM indicate that CRYAB_K92 in cooperation with S100A16 can promote the proliferation of HLF cells. Scale bar = 50 μm. G , H The results of western blot showed the regulation of CRYAB _K92 and S100A16 on the fibrosis of HLF cells and the RAGE pathway in combination with NaLA. I The lactic acid content in HLF cells treated with CRYAB _K92 site mutation, exogenous lactic acid, and S100A16 silencing. J The ATP production in HLF cells treated with CRYAB _K92 site mutation, exogenous lactic acid, and S100A16 silencing. K The ECAR in HLF cells treated with CRYAB _K92 site mutation, exogenous lactic acid, and S100A16 silencing. The quantitative data are presented as mean ± SD, with individual data points overlaid to show the distribution. * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.

Article Snippet: Recombinant S100A16 (MedChemExpress) was immobilized on a CM5 sensor chip, and serially diluted recombinant CRYAB (MedChemExpress) was injected as the analyte.

Techniques: Western Blot, Mutagenesis

Journal: Communications Biology

Article Title: CRYAB_K92 lactylation drives hypertrophy of the ligamentum flavum via an S100A16/RAGE-mediated glycolysis-fibrosis positive feedback loop

doi: 10.1038/s42003-026-10103-4

Figure Lengend Snippet: The hyperlactate state and lactylation of CRYAB_K92 promote fibrosis and hyperproliferation of ligamentum flavum cells by upregulating S100A16 and activating the RAGE pathway.

Article Snippet: Recombinant S100A16 (MedChemExpress) was immobilized on a CM5 sensor chip, and serially diluted recombinant CRYAB (MedChemExpress) was injected as the analyte.

Techniques:

Journal: Cell stem cell

Article Title: A MODULAR PLATFORM FOR DIFFERENTIATION OF HUMAN PSCS INTO ALL MAJOR ECTODERMAL LINEAGES

doi: 10.1016/j.stem.2017.08.015

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Mouse Anti-CRYAB , OriGene , {"type":"entrez-nucleotide","attrs":{"text":"CF500680","term_id":"46893706","term_text":"CF500680"}} CF500680 , RRID: AB_11125660.

Techniques: Western Blot, Virus, Recombinant, Knock-Out, Derivative Assay, Reverse Transcription, TA Cloning, Expressing, Software

Journal: Frontiers in Molecular Neuroscience

Article Title: Evaluation of the amyloid beta-GFP fusion protein as a model of amyloid beta peptides-mediated aggregation: a study of DNAJB6 chaperone

doi: 10.3389/fnmol.2015.00040

Figure Lengend Snippet: Canonical members of HSPB family prevent the aggregation of Aβ-GFP. Cells were transfected with HSPB1, HSPB5, HSPB7 or FRTTO at 1:3 ratio for 48 h. Quantification of the pellet/soluble ratio of Aβ-GFP relative to FRTTO was depicted in the chart above the blot. Values represent mean ± SE of two independent experiments. ** P < 0.01, ns = non significant.

Article Snippet: The membranes were blocked with 5% dry milk in PBS with 0.1% Tween 20 (PBST) for 1 h at room temperature and incubated overnight at 4°C with the following primary antibodies: 6E10 (1:1000 in TBST, Covance), anti V5 (1:5000 in PBST, Invitrogen), anti β-actin (1:1000 in PBST, Abcam), anti HSPB1 (1:1000 in PBST, Stress Marq Biosciences), anti HSPB5 (1:1000 in PBST, Stress Marq Biosciences) and anti HSPB7 (1:1000, Abnova).

Techniques: Transfection