rhodamine conjudated α bungarotoxin  (Alomone Labs)

Journal: Scientific Reports

Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

doi: 10.1038/s41598-022-20497-w

Figure Lengend Snippet: EDN1 and EDNRB protein expression and cell viability in 661W cultured cells under glucose-free conditions. ( a – d ) The relative amount of live cells was measured using WST-8 following 48 h treatment with DMEM/high glucose media or DMEM/glucose-free media with or without KUS121 (100 µM). ( a ) Quantitative analysis of live cells ( n = 3, for all treatments). 661W cells were cultured with high glucose [labeled ‘‘(-)’’] or treated in glucose-free media with (labeled ‘‘K121’’) or without (control, labeled ‘‘C’’) KUS121. ( b – d ) Representative images of 661W cells cultured under each condition. The black bar represents 50 µm. ( e – h ) Protein expression of EDN1 ( e , g ) and EDNRB ( f , h ) in 661W cells was analyzed by western blotting. 661W cells were cultured with high glucose media [labeled ‘‘(-)’’] or with glucose-free media with (labeled ‘‘K’’) or without (control, labeled ‘‘C’’) KUS121 for 24 h before the analysis. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. S2 b,c. Relative expression of EDN1 ( g ) and EDNRB ( h ) was shown as a ratio to actin ( n = 4, for both treatments). * p

Article Snippet: The primary antibodies were mouse anti-EDN1 antibody (1:200; Abcam) and rabbit anti-EDNRB antibody (1:100; Alomone Labs) and the secondary antibodies were biotinylated anti-mouse IgG (1:250; Vector Laboratories, Burlingame, CA, USA) for EDN1 and biotinylated anti-mouse IgG (1:250; Vector Laboratories) for EDNRB.

Techniques: Expressing, Cell Culture, Labeling, Western Blot

Journal: Scientific Reports

Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

doi: 10.1038/s41598-022-20497-w

Figure Lengend Snippet: mRNA expressions of endothelin-1 ( Edn1 ), endothelin receptor type A ( Ednra ) and endothelin receptor type B ( Ednrb ) in the mouse retina and primary retinal ganglion cells (RGCs). ( a – c ) Relative mRNA expression in neural retina. Neural retina of non-treated wild-type mice (W, n = 7) and NMDA-injected (C, n = 8), NMDA-injected-KUS121-treated (K121, n = 8) and NMDA-injected-KUS187-treated (K187, n = 8) mice were analyzed. The relative expression levels of Edn1 ( a ), Ednrb ( b ) and Ednra ( c ) mRNA were analyzed using qRT-PCR. The ratios of mRNA expression of each gene to that of glyceraldehyde-3-phosphate dehydrogenase were calculated. * p

Article Snippet: The primary antibodies were mouse anti-EDN1 antibody (1:200; Abcam) and rabbit anti-EDNRB antibody (1:100; Alomone Labs) and the secondary antibodies were biotinylated anti-mouse IgG (1:250; Vector Laboratories, Burlingame, CA, USA) for EDN1 and biotinylated anti-mouse IgG (1:250; Vector Laboratories) for EDNRB.

Techniques: Expressing, Mouse Assay, Injection, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Involvement of endothelins in neuroprotection of valosin-containing protein modulators against retinal ganglion cell damage

doi: 10.1038/s41598-022-20497-w

Figure Lengend Snippet: Protein expression of endothelin-1 (EDN1) and endothelin receptor type B (EDNRB) in retinal tissue. ( a ) The retina of non-treated wild-type (labeled ‘‘W’’), NMDA-injected (control, labeled ‘‘C’’), or NMDA-injected-KUS121-treated mice (labeled ‘‘K’’) was analyzed by western blotting using an anti-EDN1 antibody. Actin was used as a loading control. Complete scans of western blots are shown in Supplementary Fig. S2 a. ( b ) Comparison of EDN1 expression shown as ratios to actin ( n = 5, for all treatments). ** p

Article Snippet: The primary antibodies were mouse anti-EDN1 antibody (1:200; Abcam) and rabbit anti-EDNRB antibody (1:100; Alomone Labs) and the secondary antibodies were biotinylated anti-mouse IgG (1:250; Vector Laboratories, Burlingame, CA, USA) for EDN1 and biotinylated anti-mouse IgG (1:250; Vector Laboratories) for EDNRB.

Techniques: Expressing, Labeling, Injection, Mouse Assay, Western Blot

Journal: bioRxiv

Article Title: Mutation in the TRKB cholesterol recognition site that blocks antidepressant binding does not influence the basal or BDNF-stimulated activation of TRKB

doi: 10.1101/2022.08.26.505413

Figure Lengend Snippet: (A) The sequence of TRKB transmembrane (TM) domain from rat (UniProt: Q63604, residues 430-453) was mutated at Y433 residue (Y433F) or the entire motif was substituted by the rat sequence of TRKA TM (P35739, residues 419-442). (B) The interaction between biotinylated BDNF (bBDNF) and TRKB is not affected by the TRKB.Y433F mutation or in TRKB/TRKA.TM constructs. (C) TRKB.Y433F does not prevent the BDNF-induced dimerization of TRKB. The constructs used allow the formation of TRKB.wt homodimer or TRKB.wt/Y433F heterodimer. (D , E) Analysis of TRKB phosphorylation shows that, cortical cultures from TRKB.Y433F heterozygous mouse embryos respond to BDNF similarly to the the TRKB.wt littermates, regardless of the tyrosine residue tested ( D : Y515, E : Y816).

Article Snippet: The plates were then washed 3x with PBS buffer, and various concentrations of biotinylated BDNF (Alomone Labs, cat#B-250, 0.1 – 100 pM) was added for 1h at RT.

Techniques: Sequencing, Mutagenesis, Construct