leptomycin b  (Santa Cruz Biotechnology)

Journal: bioRxiv

Article Title: NBR1 shuttles between the cytoplasm and nucleus and is essential for nuclear p62 body formation

doi: 10.64898/2026.03.10.710728

Figure Lengend Snippet: (A) HeLa cells stained for endogenous NBR1 (green) and the nuclear envelope marker lamin B1 (white). White arrows indicate nuclear NBR1 puncta. Scale bars equal to 10 µm. (B) HEK293-, HeLa-, and U2OS cells treated with leptomycin B (LMB, 3 h) or not (control). Cells were stained for NBR1 (green) and lamin B1 (white). Scale bars equal to 10 µm. (C) Quantification of number of NBR1 puncta per nucleus in B (n=50 cells). (D) Quantification of mean ± SD area of NBR1 puncta (µm²) in B (n=50 cells). Statistical comparison by One-way ANOVA. ****p < 0.0001. (E) HeLa Flp-In T-REx EGFP-NBR1 cells were treated with tetracycline (24h) to induce expression of EGFP-NBR1 (green) and subsequently subjected to LMB (3h). Cells were stained for lamin B1 (white). (F) Nuclear NBR1 fluorescence in E plotted as a function of total cellular NBR1 fluorescence for individual cells (n=50 cells).

Article Snippet: The following reagents were used: 1,6-hexanediol (Sigma-Aldrich #240117), puromycin (Sigma-Aldrich, P8833), DMEM (Sigma-Aldrich #D6429), Penicillin-Streptomycin (Sigma-Aldrich #P4333), FBS (Sigma-Aldrich, #F7524), hygromycin (Thermo Fisher Scientific #10687010), blasticidin (Thermo Fisher Scientific #A1113903), TransIT®-LT1 Transfection Reagent (Mirus #MIR 2304), Leptomycin B (Santa Cruz #sc-358688), tetracycline hydrochloride (Sigma-Aldrich #T7660).

Techniques: Staining, Marker, Control, Comparison, Expressing, Fluorescence

Journal: Cellular and Molecular Neurobiology

Article Title: Nitric Oxide Donor Alleviates Cardiac Arrest Induced Blood Brain Barrier Injury by Inhibiting HMGB1-ATG5 Mediated Endothelial Autophagy

doi: 10.1007/s10571-026-01706-w

Figure Lengend Snippet: Inhibition of HMGB1 nucleocytoplasmic translocation attenuates autophagy and improves endothelial function in bEnd.3 cells. A Representative immunoblot images and B quantitation of nuclear HMGB1, cytoplasmic HMGB1, LC3B II, LAMP2, occludin and ZO-1 expression in bEnd.3 cells from Control, OGD/R and OGD/R + Leptomycin B groups. Histone H3 and β-actin were used as the protein loading control. The images of western blotting data derived from triplicate blots conducted as three independent experiments. C Representative immunofluorescence images of HMGB1 localization in bEnd.3 cells from Control, OGD/R and OGD/R + Leptomycin B groups. HMGB1 (green), nuclei stained with DAPI (blue). Scale bar = 50 μm. D Quantification of nuclear and cytoplasmic HMGB1 signals. Nuclear HMGB1 was defined as HMGB1 signal overlapping with DAPI, and cytoplasmic HMGB1 as non-overlapping HMGB1 signal. E Representative images of fluorescent LC3 dots are shown. F Mean number of autophagosomes and autolysosomes per cell. Scale bar = 10 μm. G The diffusion rates of FITC-dextran (40 kDa). H Representative images of transwell migration assay and I quantitative analysis of migration cells. Scale bar = 100 μm. J Representative images of tube formation assay and K quantitative analysis of total tube length. Scale bar = 100 μm

Article Snippet: Leptomycin B (a selective inhibitor of CRM1-dependent nuclear export, used to block HMGB1 nuclear-to-cytoplasmic translocation) was purchased from MedChemExpress (HY-16909, China).

Techniques: Inhibition, Translocation Assay, Western Blot, Quantitation Assay, Expressing, Control, Derivative Assay, Immunofluorescence, Staining, Diffusion-based Assay, Transwell Migration Assay, Migration, Tube Formation Assay

Journal: Cellular and Molecular Neurobiology

Article Title: Nitric Oxide Donor Alleviates Cardiac Arrest Induced Blood Brain Barrier Injury by Inhibiting HMGB1-ATG5 Mediated Endothelial Autophagy

doi: 10.1007/s10571-026-01706-w

Figure Lengend Snippet: HMGB1 regulates autophagy in endothelial cells through interacting with ATG5. A PPI network functional enrichment analysis of HMGB1 and ATG5. B Molecular docking model showing the predicted interaction interface between HMGB1 (yellow) and ATG5 (blue). The surface structure of the docking complex is displayed on the left. The enlarged panel on the right shows the predicted binding interface and key interacting residues. Hydrogen bonds are indicated by yellow dashed lines. The calculated binding free energy (ΔG) was − 7.9 kcal/mol. C Co-IP analysis of the interaction between HMGB1 and ATG5 in bEnd.3 cells under OGD/R condition. Cell lysates were immunoprecipitated with anti-HMGB1 or anti-ATG5 antibodies, followed by immunoblotting with antibodies against ATG5 or HMGB1. IgG was used as a negative control. D Co-IP of HMGB1 and ATG5 in bEnd.3 cells under different conditions. Cell lysates from Control, OGD/R and OGD/R + Leptomycin B groups were immunoprecipitated with anti-HMGB1 antibody and blotted for ATG5 and HMGB1. IgG was used as a negative control

Article Snippet: Leptomycin B (a selective inhibitor of CRM1-dependent nuclear export, used to block HMGB1 nuclear-to-cytoplasmic translocation) was purchased from MedChemExpress (HY-16909, China).

Techniques: Functional Assay, Binding Assay, Co-Immunoprecipitation Assay, Immunoprecipitation, Western Blot, Negative Control, Control