Journal: Scientific Reports

Article Title: fMRI indicates cortical activation through TRPV1 modulation during acute gouty attacks

doi: 10.1038/s41598-019-48656-6

Figure Lengend Snippet: Cortical TRPV1 expression was enhanced by gouty arthritis. ( A ) ROI selection. ( B ) Cortical TRPV1 expression on the MSU-affected and control sides. ( C ) TRPV1 immunoreactivity intensity. ( D ) TRPV1 immunoreactivity coverage. ( E ) Immunoprecipitation of beta-tubulin with TRPV1 immunoblotting, which demonstrates a strong interaction between TRPV1 and neuronal filament proteins. The cropped images of the gels are used in the figure, and images of the full-length gels are presented in Supplementary Fig. S2. (***Indicates statistically significant difference with p < 0.001 after ANOVA). Scale bar = 50 µm.

Article Snippet: The primary antibodies used were: anti-TRPV1 (1:2000; PC420; Merck Millipore, Darmstadt, Germany); anti-COX-2 (1:1000; Thermo Fisher Scientific); β-tubulin (1:2000; MCA2703; Bio-Rad Laboratories); β-actin (1:4000; sc-47778; Santa Cruz Biotechnology).

Techniques: Expressing, Selection, Control, Immunoprecipitation, Western Blot

Journal: Scientific Reports

Article Title: The Role of Transthyretin in Oligodendrocyte Development

doi: 10.1038/s41598-020-60699-8

Figure Lengend Snippet: TTR influences OPC differentiation. Representative images of neurospheres in 3D culture, isolated from SVZ derived NSCs of P21 ( A ) wild type and ( B ) TTR null mice. Scale bar 5 mm. ( C ) Quantitation of the potency of the NSC colonies isolated from the SVZ of P21 mice by neural colony-forming cell assays. There was a significant increase in the number of colonies with an average diameter ≥ 2 mm that were isolated from the SVZ of P21 TTR null mice when compared with wild type at the same age. Increases corresponded with a significant decrease in the number of colonies with an average diameter < 2 mm that were isolated from TTR null mouse SVZ compared to wild type. The colonies ≥ 2 mm in diameter are “NSC derived” and have self-renewal and multi-potential capabilities. Colonies < 2 mm diameter are “progenitor derived”. All data were expressed as the mean ± SEM. Statistical comparisons were performed using a one-way ANOVA with post hoc analysis using Sidak’s test as appropriate (data derived from n = 4 independent experiments per genotype and P < 0.05 was considered statistically significant). ( D ) Characterization of NSCs isolated from the SVZ of P21 TTR null mice and differentiated into the three neural lineages: oligodendrocytes, astrocytes and neurons. Nuclei were stained with DAPI; oligodendrocytes were stained with anti-Olig2; neurons were stained with anti-β-III-tubulin; astrocytes were stained with anti-GFAP. Lack of TTR promotes NSCs to differentiate into glial precursor cells. Differentiation assay for NSCs isolated from the SVZ of P21 TTR null mice showed a greater proportion of cells differentiating into a glial lineage, whereas the equivalent cells from wild type mice had a greater proportion differentiating into a neuronal lineage. All data were expressed as the mean ± SEM. Statistical comparisons were performed using a one-way ANOVA with post hoc analysis using Sidak’s test as appropriate (n = 3 independent experiments and P < 0.05 was considered statistically significant). (Modified from ).

Article Snippet: Cell pellets were collected and incubated with primary antibodies diluted in blocking solution: mouse anti-Olig2 (1:200; Merck Millipore, MAPN50), mouse anti-beta III tubulin (1:150; Merck Millipore, MAB1637), mouse anti-GFAP (1:200; Merck Millipore, MAB360), rabbit anti-TTR (1:300;; ABBIOTC, 250892) and incubated overnight at 4 °C.

Techniques: Isolation, Derivative Assay, Quantitation Assay, Staining, Differentiation Assay, Modification

Journal: PLoS ONE

Article Title: Beta-Actin Is Required for Proper Mouse Neural Crest Ontogeny

doi: 10.1371/journal.pone.0085608

Figure Lengend Snippet: (A) Neurofilament expression in Actb +/+ and Actb −/− embryos. Aberrations of neurofilament expression in Actb −/− embryos are visible at the level of all nerves. (B) Beta-III tubulin expression in Actb +/+ and Actb −/− embryos. Aberrations of beta-III tubulin expression in Actb −/− embryos are visible at the level of IX and X nerves and the dorsal root ganglia. (C) Western Blot analysis showing presence of GFP expression (driven by the endogenous promotor) and the lack of beta-actin expression in Actb −/− embryos. V: trigeminal nerve; VII: facial nerve; IX: glossopharyngeal nerve; X: vagus nerve; max.b: maxillary branch; man.b: mandibular branch; op.b: ophthalmic branch; drg: dorsal root ganglia. Panels depicting Actb +/+ and Actb −/− embryos are of the same magnification.

Article Snippet: Antibodies used in this study are mouse anti-neurofilament mAb (clone 2H3) from Developmental Studies Hybridoma Bank; mouse anti-beta-III tubulin mAb (clone5G8) from Promega; anti-digoxigenin-AP Ab from Roche; mouse anti-p75 NGF receptor mAb, rabbit anti-p75 NGF receptor pAb, mouse anti-Histone H3 (phospho S10) mAb, rabbit anti-vimentin mAb, all from Abcam; rat anti-E-cadherin mAb (clone DECMA-1) from Sigma-Aldrich; mouse anti-N-cadherin mAb (clone 3B9), mouse anti-cadherin-11 mAb (clone 5B2H5), anti-mouse Alexa Fluor 488, anti-rabbit Alexa Fluor 488, anti-rabbit Alexa Fluor 594, anti-rat Alexa Fluor 594, anti-mouse Alexa Fluor 647, anti-rabbit Alexa Fluor 647 all from Invitrogen.

Techniques: Expressing, Western Blot

Journal: iScience

Article Title: Sorafenib induces cachexia by impeding transcriptional signaling of the SET1/MLL complex on muscle-specific genes

doi: 10.1016/j.isci.2024.110913

Figure Lengend Snippet: Effect of TKIs on sarcomere organization (A) Panel shows confocal microscopy images of TKI-treated myotubes immunostained against alpha-actinin (green) and nucleus (DAPI – blue). Images were captured at 4× magnification over 60× oil immersion objective. Scale bar = 10 μm. See also Figure S1 . The images are the representative of at least three biological experiments (N). The concentrations used for Nilo was 0.1 μM, and 20 μM for Ima and Sor. 0.1% DMSO was used as vehicle control. (B–D) Western blot showing protein expression of MyHC-IId in myotubes treated with (B) Nilo, (C) Ima, and (D) Sor. (E–G) Western blot showing protein expression of MyHC-IIa in myotubes treated with (E) Nilo, (F) Ima, and (G) Sor. β-tubulin was used as the loading control. The blots are representative of three biological experiments (N). All concentrations are in μM. See also Figure S2 .

Article Snippet: Mouse monoclonal anti-beta tubulin , Biocat , Cat#AKR-009.

Techniques: Confocal Microscopy, Control, Western Blot, Expressing

Journal: iScience

Article Title: Sorafenib induces cachexia by impeding transcriptional signaling of the SET1/MLL complex on muscle-specific genes

doi: 10.1016/j.isci.2024.110913

Figure Lengend Snippet:

Article Snippet: Mouse monoclonal anti-beta tubulin , Biocat , Cat#AKR-009.

Techniques: Recombinant, Isolation, SYBR Green Assay, Protease Inhibitor, Control, Software, Real-time Polymerase Chain Reaction