Journal: Nature Communications

Article Title: Alternative antigen retention by a gp96-fusion approach induces long-lasting and broad immunity in mice

doi: 10.1038/s41467-025-67092-x

Figure Lengend Snippet: A Large-scale analysis of the key binding regions of antigens with GP96 using AlphaFold3 ( n = 22). B The possible structural pattern diagram of the combination of RBD and GP96 modeled by AlphaFold3. C The schematic representation of RBD-GP96-Fusion (monomer). RBDs (prototype strain) are truncated at the C-terminal residue G545 and connected to GP96 protein N-terminal residue D22 to construct the fusion protein (SP, signal peptide). The structure of the fusion protein was predicted based on AlphaFold3 (yellow RBM motif, hACE2 binding domain). D , E Molecular dynamics of GP96-dimer and RBD-GP96-dimer predicted by AlphaFold2-multimer were performed using AMBER. D The RMSD was analyzed. The red line represents RBD-GP96-dimer and the black line represents GP96-dimer. E The last frame of the molecular dynamics operation of the dimer conformations is captured for display. F Analytical gel filtration of the RBD-GP96-Fusion protein was performed with Superdex 200 Increase 10/300 GL. The resulting 280-nm absorbance curve is shown. The reduced SDS-PAGE migration profile of the pooled sample is shown. The data are representative of two independent experiments with similar results. G Ultracentrifugation sedimentation profiles of RBD-GP96-Fusion. H Transmission electron microscopy of RBD-GP96-Fusion and GP96 protein. The red frames represent typical transmission electron microscopy (TEM) structures. Below is a 3D cartoon sketch of the RBD-GP96-Fusion (surface, cone). I Analysis of GP96 and RBD-GP96-Fusion dimerization models.

Article Snippet: D , E Molecular dynamics of GP96-dimer and RBD-GP96-dimer predicted by AlphaFold2-multimer were performed using AMBER.

Techniques: Binding Assay, Residue, Construct, Filtration, SDS Page, Migration, Sedimentation, Transmission Assay, Electron Microscopy

Journal: The Journal of Biological Chemistry

Article Title: The ER-associated degradation adaptor SEL1L is dispensable for ER homeostasis and the differentiation of spermatogenic cells

doi: 10.1016/j.jbc.2025.110283

Figure Lengend Snippet: ER homeostasis is not affected by male germ cell-specific Sel1L ablation. A and B , RT-PCR analysis of Xbp1 splicing ( A ) and qPCR analysis ( B ) in testes from 6-week-old WT Stra8 , HET Stra8 , and KO Stra8 littermates. In ( A ), u and s denote unspliced and spliced Xbp1 mRNA, respectively. Percentage of Xbp1 splicing is quantitated below the images. Male germ GC-4 cells treated with 2 μg/ml tunicamycin (Tuni) for 4 h serve as a positive control. N = 4 in A, n = 5 in B. C and D , Western blot analysis of testes from 6-week-old WT Stra8 , HET Stra8 , and KO Stra8 littermates, with quantitation normalized to HSP90 shown in ( D ). GC-4 cells treated with 2 μg/ml tunicamycin for 4 h serve as a positive control. N = 4 per cohort. E , immunofluorescent staining of GRP94 in testis sections from 6-week-old HET Stra8 and KO Stra8 littermates. Representative images from n = 4 per cohort. F , Western blot analysis of testes from 6-week-old WT Stra8 , HET Stra8 , and KO Stra8 littermates. GC-4 cells treated with 2 μg/ml tunicamycin for 24 h serve as a positive control. G , TUNEL staining ( green ) with nuclei stained with DAPI in testes from 6-week-old HET Stra8 and KO Stra8 littermates. White arrowheads, TUNEL-positive cells. The cell death index represents the percentage of seminiferous tubules containing at least one apoptotic cell, quantified from low-magnification whole testis sections of n = 8 mice per cohort. Values, mean ± SD. ∗∗∗, p < 0.001; ns or not labeled, not significant by one-way ANOVA with Tukey multiple comparison tests in ( A , B and D ), and by two-tailed Student’s t test in ( G ).

Article Snippet: The following antibodies were used in this study: SEL1L (Abclonal customized) , HRD1 (Abclonal customized E15102 , used for mouse tissues) , HRD1 (proteintech 13473-1-AP, used for HEK293T cell), OS9 (Abcam ab109510), IRE1α (CST 14C10), BiP (CST C50B12), GRP94 (Novus NBP2-42379), calmegin (Proteintech 12629-1-AP), PERK (CST 3192), ATF4 (CST 11815), calnexin (Enzo ADI-SPA-860-D), PDI (Enzo ADI-SPA-890), CD147 (Proteintech 11989-1-AP), caspase-3 (CST 9662), and HSP90 (Santa Cruz sc7947).

Techniques: Reverse Transcription Polymerase Chain Reaction, Positive Control, Western Blot, Quantitation Assay, Staining, TUNEL Assay, Labeling, Comparison, Two Tailed Test