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: Stem Cell Research & Therapy

Article Title: SERCA-mediated endoplasmic reticulum stress facilitates hematopoietic stem cell mobilization

doi: 10.1186/s13287-025-04345-y

Figure Lengend Snippet: Compounds and abbreviation

Article Snippet: KN-93 Phosphate , KN93 , Selleck , S7423.

Techniques:

Journal: Stem Cell Research & Therapy

Article Title: SERCA-mediated endoplasmic reticulum stress facilitates hematopoietic stem cell mobilization

doi: 10.1186/s13287-025-04345-y

Figure Lengend Snippet: ER Stress triggers calcium signaling pathway to regulate CXCR4 expression. ( A-B ) Downregulation of CXCR4 surface expression in SERCA2/3 knockdown Jurkat cells ( n = 3). ( C ) The reduced CXCR4 expression could not be restored by the SERCA agonist (CDN1163) in SERCA2/3 knockdown Jurkat cells ( n = 3). ( D ) Flow cytometric analysis of intracellular calcium transients in BHQ-treated Jurkat cells ( n = 3). ( E ) Western blot analysis of p-CaMKII and CaMKII protein levels following BHQ treatment. Full-length blots are presented in Supplementary Fig. . ( F-G ) Suppression of CXCR4 expression in Jurkat cells treated with BD (250 µM for 1.5 h) and KN93 (100 µM for 5 h), similar to the effect of BHQ ( n = 3). ( H-I ) Suppression of CXCR4 expression in mouse LSKs and HSCs by CaMKII inhibitors ( n = 3)

Article Snippet: KN-93 Phosphate , KN93 , Selleck , S7423.

Techniques: Expressing, Knockdown, Western Blot

Journal: Stem Cell Research & Therapy

Article Title: SERCA-mediated endoplasmic reticulum stress facilitates hematopoietic stem cell mobilization

doi: 10.1186/s13287-025-04345-y

Figure Lengend Snippet: CaMKII inhibitors promote HSPC mobilization in vivo. ( A , D ) CFU counts per milliliter of PB following treatment with BD ( n = 9) or KN93 ( n = 6). ( B-C , E-F ) Absolute numbers of LSKs and HSCs per milliliter of PB, as determined by flow cytometry, following treatment with BD ( n = 9) or KN93 ( n = 6). ( G-L ) Enhanced HSPC mobilization when G-CSF is combined with BD ( n = 6) or KN93 ( n = 5). n indicates the number of mice used per group

Article Snippet: KN-93 Phosphate , KN93 , Selleck , S7423.

Techniques: In Vivo, Flow Cytometry

Journal: Stem Cell Research & Therapy

Article Title: SERCA-mediated endoplasmic reticulum stress facilitates hematopoietic stem cell mobilization

doi: 10.1186/s13287-025-04345-y

Figure Lengend Snippet: STAT3 and CXCR4 phosphorylation in Jurkat cells following BHQ or KN93 treatment. ( A-E) Western blot analysis of protein levels of STAT3, pSTAT3 (Y705), CXCR4, and pCXCR4 (S339) in Jurkat cells treated with BHQ for 0.5, 1.5, and 5 h. ( F-J ) Western blot analysis of protein levels of STAT3, pSTAT3 (Y705), CXCR4, and pCXCR4 (S339) in Jurkat cells after KN93 stimulation for 0.5, 1.5, and 5 h. Full-length blots are presented in Supplementary Fig.

Article Snippet: KN-93 Phosphate , KN93 , Selleck , S7423.

Techniques: Phospho-proteomics, Western Blot

Journal: Cell reports

Article Title: LTP expression mediated by autonomous activity of GluN2B-bound CaMKII

doi: 10.1016/j.celrep.2024.114866

Figure Lengend Snippet: LTP was induced by 2× HFS and measured by the fEPSP slopes at the CA3 to CA synapses in hippocampal slices from wild-type (WT) mice. Drug (10 μM unless noted otherwise) was added as indicated. Data show mean ± SEM. (A) When added 1 min after LTP induction, the ATP-competitive inhibitors AS397 and ruxolitinib (Ruxo) both reduced fEPSP slopes back to baseline within 60 min ( n = 7, 6, and 6 slices for control without drug, Ruxo, and AS397, respectively). (B) Significant reduction of LTP during the last 5 min of recording was observed for both AS397 and Ruxo (** p < 0.005 and *** p < 0.001 by Bonferroni multiple comparison test following one-way ANOVA). (C) When added 1 min after LTP induction, the peptide inhibitor tatCN21 (5 μM), but not the CaM-competitive inhibitor KN93, reduced fEPSP slopes back to baseline within 60 min ( n = 6 for KN93 and tatCN21). (D) During the last 5 min of recordings, only tatCN21, but not KN93, caused a significant reduction of fEPSP slope compared to the no-drug control, as also shown in the other images, and tatCN21 also caused significant reduction compared to KN93 (*** p < 0.001 and * p < 0.05 by one-way ANOVA followed by Bonferroni multiple comparison test). (E) The effect of different inhibitors (10 μM) on the autonomous activity of GluN2B-bound CaMKII was tested in vitro as illustrated in the schematic on the right. CaMKII binding was induced by Ca 2+ /CaM, which was then washed out before measuring the Ca 2+ /CaM-independent autonomous activity of the bound kinase. Negative control (−) contained no kinase and positive control (+) no inhibitor. All ATP-competitive inhibitors (AS105, AS283, and AS397) and tatCN21 completely blocked activity. KN93 had no effect on activity, and AIP reduced activity but did not block it completely. (F) When added 15 min after LTP induction, the ATP-competitive inhibitor AS397 had no apparent effect on LTP maintenance ( n = 8 slices). (G) During the last 5 min of recording, addition of ATP at 15 min after LTP induction had no significant effect on fEPSP slope compared to the control without the drug, which is also shown in the other images.

Article Snippet: KN93 , Tocris , Catalog #: 5215.

Techniques: Control, Comparison, Activity Assay, In Vitro, Binding Assay, Negative Control, Positive Control, Blocking Assay

Journal: Cell reports

Article Title: LTP expression mediated by autonomous activity of GluN2B-bound CaMKII

doi: 10.1016/j.celrep.2024.114866

Figure Lengend Snippet: Data show mean ± SEM. (A) In slices from mice with the GluN2B ΔCaMKII mutant that prevents CaMKII binding to GluN2B, neither 5 μM tatCN21 nor 10 μM KN93 (two CaMKII inhibitors that block LTP induction in wild-type mice) reduced LTP compared to control without drug ( n = 9,8, and 7 slices for control, tatCN21, and KN93, respectively), showing that the LTP seen in the mutant mice was enabled by compensatory mechanisms and different from LTP in wild type. Drug was added 15 min before and washed out 5 min after LTP induction, as indicated. LTP was induced by 2× HFS and measured by the fEPSP slopes at the CA3 to CA synapses. (B) During the last 5 min of recording, no statistical difference was detected in the fEPSP slopes in the tatCN21, KN93, or control condition (one-way ANOVA). (C) Co-condensation of CaMKII wild type with the cytoplasmic C-tail of GluN2B in HEK cells is triggered by a Ca 2+ stimulus with ionomycin and then maintained after chelating Ca 2+ with EGTA (even if at a reduced level). For a CaMKII T286A mutant, Ca 2+ triggers at least as much co-condensation, but these condensates are then fully reversed after chelating Ca 2+ with EGTA. Scale bar: 10 μm.

Article Snippet: KN93 , Tocris , Catalog #: 5215.

Techniques: Mutagenesis, Binding Assay, Blocking Assay, Control

Journal: Cell reports

Article Title: LTP expression mediated by autonomous activity of GluN2B-bound CaMKII

doi: 10.1016/j.celrep.2024.114866

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: KN93 , Tocris , Catalog #: 5215.

Techniques: Virus, Recombinant, Saline, Protease Inhibitor, Software, Imaging, Modification