myr aip  (Tocris)

Journal: bioRxiv

Article Title: Direct interaction of ribosomes with postsynaptic proteins gives rise to a privileged local synaptic translatome

doi: 10.64898/2026.02.27.708433

Figure Lengend Snippet: ( A ) Schematic of XL-MS workflow. ( B ) XL-based interactome of synaptic ribosomes. Synaptic annotation is based on SynGO (dataset version 20231201). ( C ) Protein domain-level crosslink map between RPL35 and CaMKIIα. Domain classification based on . ( D ) Structural model of CaMKIIα binding to the ribosome. Most CaMKIIα–RPL35 crosslinks satisfy distance constraints (green), with remaining links accommodated by CaMKIIα linker flexibility (yellow). ( E ) Detection of newly-synthesized proteins in dendrites and synapses from control (untreated) neurons or neurons treated with 10 μM KN-93 or myr-AIP for 30 min. Nascent proteins were labeled with puromycin (5 min) in the absence or presence of the protein synthesis inhibitor anisomycin (see Methods). Scale bar, 5 µm. ( F ) Quantification of puromycin signal in postsynaptic regions of the apical dendritic arbor from control and treated neurons. Error bars show mean ± SEM. **p<0.01; ****p<0.0001, Brown-Forsythe and Welch’s ANOVA, Dunnett’s multiple comparisons test; n=58-64 neurons from 3 independent cultures.

Article Snippet: DIV 18-19 hippocampal neurons were treated with either 10 μM KN-93 (Tocris, #5215) or 10 μM myr-AIP (Tocris, #5959) for 30 min at 37°C, or left untreated.

Techniques: Structural Proteomics, Binding Assay, Synthesized, Control, Labeling

Journal: Cells

Article Title: MLKL and CaMKII Are Involved in RIPK3-Mediated Smooth Muscle Cell Necroptosis

doi: 10.3390/cells10092397

Figure Lengend Snippet: CaMKII is partially required for SMC necroptosis. ( A – D ) MOVAS cells were pretreated with indicated doses of CaMKII inhibitor myristoylated autocamtide-2-related inhibitory peptide (Myr-AIP) for 1 h, followed by 6 h of treatment with 30 ng/mL TNFα plus 60 µM zVAD. ( A ) Cells were lysed in RIPA buffer and subjected to Western blotting analysis with the indicated antibodies. ( C ) Cells were stained with 7-AAD and Annexin V, and subjected to flow cytometry analysis. Necrotic cells were identified as 7-AAD + Annexin V + . ( B ) was quantification of ( A ). ( D ) was quantification of ( C ). ( E ) MOVAS cells were transfected with siRNAs against Camk2d for 48 h. Cells were lysed in RIPA buffer and subjected to Western blotting analysis with the indicated antibodies. ( F , G ) Quantification of ( E ). ( H ) MOVAS cells were transfected with siRNAs against Camk2d for 48 h, then treated with 30 ng/mL TNFα plus 60 µM zVAD for 6 h. Cells were then stained with 7-AAD and Annexin V, and subjected to flow cytometry analysis. Necrotic cells were identified as 7-AAD + Annexin V + . ( I ) Quantification of ( H ). Data were presented as mean ± SD of at least three independent experiments. One-way ANOVA was performed in ( B , D , F , G , I ). * p < 0.05, ** p < 0.01, *** p < 0.001 compared with TNFα plus zVAD treated group ( B , D , I ) or scrambled siRNA-treated group ( F , G ).

Article Snippet: Myristoylated autocamtide-2-related inhibitory peptide (Myr-AIP) was purchased from Enzo (Farmingdale, NY, USA.

Techniques: Western Blot, Staining, Flow Cytometry, Transfection