Journal: Redox Biology

Article Title: PHB2 ameliorates ferroptosis and aortic aneurysm/dissection through NEDD4L-dependent ubiquitination of NCOA4

doi: 10.1016/j.redox.2026.104114

Figure Lengend Snippet: PHB2-evoked NCOA4 degradation through ubiquitination. (A and B). Representative Western blotting images and quantification of NCOA4 expression in VSMCs with or without PHB2 expression (n = 6 per group, p < 0.0001). C. Quantification of relative mRNA levels of NCOA4 in VSMCs with or without PHB2 overexpression (n = 6 per group). D. Representative Western blotting images depicting the effect of proteasome inhibitor (MG132) and lysosomal inhibitors (Pepstatin A + E64d) on NCOA4 protein levels in VSMCs with or without PHB2 overexpression. (E and F) Representative Western blotting images and quantification of NCOA4 protein stability in CHX-chase assays. G. Representative Co-IP images showing the ubiquitination of NCOA4 with or without PHB2 overexpression. H. Representative Co-IP images showing the ubiquitination of NCOA4 with or without PHB2 silencing. Mean ± SEM. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, ns, no significance.

Article Snippet: To determine whether protein degradation was mediated by the ubiquitin proteasome pathway, cells were treated with the proteasome inhibitor MG132 (10 μM; MedChemExpress) for 6 h before harvest.

Techniques: Ubiquitin Proteomics, Western Blot, Expressing, Over Expression, Co-Immunoprecipitation Assay

Journal: Poultry Science

Article Title: RACK1 inhibits fowl adenovirus serotype 4 replication by targeting the viral protein Hexon for ubiquitin-proteasome degradation

doi: 10.1016/j.psj.2026.106627

Figure Lengend Snippet: Interaction with RACK1 targets Hexon for degradation through the ubiquitin-proteasome pathway. ( A ) LMH cells were co-transfected with Flag-Hexon (2 µg) and Myc-RACK1 (2 µg) for different time (24 and 48 h). After transfection, the cell lysates were harvested and examined by Western blotting. ( B ) LMH cells were co-transfected with Flag-Hexon and Myc-RACK1 (0.4 or 2 µg) for 24 h. After transfection, the cell lysates were harvested and examined by Western blotting. ( C ) The relative expression levels of Flag-Hexon in panel A. ( D ) The relative expression levels of Flag-Hexon in panel B. ( E ) LMH cells were co-transfected with Flag-Hexon and Myc-RACK1 or Flag-Hexon and empty vector. At 24 h post transfection, cells were treated with cycloheximide (CHX, 50 μg/mL) at different time points (0, 2, 4, 6, and 8 h) before being harvested. Cell lysates were prepared and examined by Western blotting using the indicated antibodies. (F ) The relative expression levels of Flag-Hexon in panel E. ( G ) LMH cells were pretreated with DMSO, Z-VAD-FMK, NH 4 Cl for 2 h, followed by co-transfection with Flag-Hexon and Myc-RACK1 for 24 h in the presence of the respective inhibitors. Cell lysates were prepared and examined with Western blotting using the indicated antibodies. For MG132 (10 μM), cells were not pretreated; instead, this inhibitor was added to the culture medium 12 h prior to sample collection. ( H ) The relative expression levels of Flag-Hexon in panel G. Data are representative of three independent experiments and presented as means ± SD. ***, P < 0.001; **, P < 0.01; *, P < 0.05; ns, P > 0.05.

Article Snippet: Dimethyl sulfoxide (DMSO), Z-VAD-FMK, MG132, BafA1, NH 4 Cl were purchased from MedChemExpress (MCE, China).

Techniques: Ubiquitin Proteomics, Transfection, Western Blot, Expressing, Plasmid Preparation, Cotransfection