p53  (Proteintech)

Journal: Regenerative Therapy

Article Title: Hypoxia-preconditioned bone marrow mesenchymal stem cell-derived exosomes ameliorate knee osteoarthritis by promoting cartilage regeneration and alleviating pain in rats

doi: 10.1016/j.reth.2025.101049

Figure Lengend Snippet: Effects of Hypo-BMSCs-Exos on IL-1β-induced chondrocyte senescence. A: Immunofluorescence staining revealed changes in p16, p21, and p53 in IL-1β-induced chondrocytes following intervention with Norm-BMSCs-Exos and Hypo-BMSCs-Exos. B: Quantitative analysis of immunofluorescence staining. C: Western blot detection of p16, p21, and p53 expression in chondrocytes. D: Statistical analysis of gray values and relative values for corresponding protein bands. E: SA-β-gal activity in chondrocytes across different groups assessed using SA-β-gal staining kit, bar = 75 μm. F: Counting and statistical analysis of SA-β-gal positive cells in chondrocytes across groups. G: Flow cytometry analysis of mean fluorescence intensity for reactive oxygen species (ROS). H: Statistical analysis of ROS expression levels across groups. Results are presented as Mean ± SD. Intergroup statistical significance is indicated as: ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 (n = 3).

Article Snippet: The cells were then washed three times with PBS, with each wash lasting 5 min. After blocking with bovine serum albumin (BSA) for 20 min at room temperature and subsequent washing with PBS, the cells were incubated overnight at 4 °C with primary antibodies against iNOS (Servicebio, China), COX2 (Servicebio, China), Collagen II (Bisso, China), aggrecan (Bisso, China), ADAMTS-5 (Bisso, China), MMP-13 (Affinity, USA), p16 (Abcam, USA), p21 (Affinity, USA), and p53 (Proteintech, USA).

Techniques: Immunofluorescence, Staining, Western Blot, Expressing, Activity Assay, Flow Cytometry, Fluorescence

Journal: International Dental Journal

Article Title: FBXO5 alleviates apical periodontitis by facilitating TP53 protein degradation

doi: 10.1016/j.identj.2025.103948

Figure Lengend Snippet: FBXO5 promotes TP53 degradation through ubiquitination. (A) UbiBrowser was used to identify downstream targets of FBXO5. Circular nodes represented proteins predicted as FBXO5 substrates by UbiBrowser. Orange lines indicated domain-pair evidence, purple lines indicated GO term-pair evidence, green lines indicated E3-recognising motif evidence, and black lines indicated network-loop evidence. TP53 was highlighted with a red circle as a predicted substrate of FBXO5. (B-C) RT-qPCR and western blotting were employed to assess the mRNA and protein expression levels of TP53 in hSCAPs transfected with shNC or shFBXO5 (effect size = 0.57 for B, 5.87 for C). (D) Co-IP was performed to analyse the interaction between FBXO5 and TP53. (E) The protein level of TP53 was analysed by Western blotting in hSCAPs transfected with shNC or shFBXO5, followed by CHX treatment at various time points (0 hours, 4 hours, 8 hours, 12 hours). (F) Ubiquitination of TP53 was evaluated using Co-IP analysis after FBXO5 knockdown. Data were presented as mean ± SD. ns, no significance. Exact P values were indicated in the figure, P < .05 was considered statistically significant (Unpaired t-test for B-C).

Article Snippet: After blocking with 5% skim milk, the membranes were incubated overnight at 4°C with anti-FBXO5 antibody (1:1000, ab187144, Abcam), anti-TP53 antibody (1:3000, 10442-1-AP, Proteintech, Wuhan, China), anti-runt-related transcription factor 2 (RUNX2) antibody (1:2000, 20700-1-AP, Proteintech), anti-DLX5 antibody (1:1000, 10592-1-AP, Proteintech), anti-osterix (OSX) antibody (1:1000, 28694-1-AP, Proteintech), anti-osteocalcin (OCN) antibody (1:1000, ab133612, Abcam), or anti-GAPDH antibody (1:1000, ab9485, Abcam).

Techniques: Ubiquitin Proteomics, Quantitative RT-PCR, Western Blot, Expressing, Transfection, Co-Immunoprecipitation Assay, Knockdown

Journal: International Dental Journal

Article Title: FBXO5 alleviates apical periodontitis by facilitating TP53 protein degradation

doi: 10.1016/j.identj.2025.103948

Figure Lengend Snippet: FBXO5 enhances hSCAPs proliferation by suppressing TP53 expression. (A) The hSCAPs transfected with oeNC, oeFBXO5, or oeTP53 were incubated in osteogenic induction medium for 14 days. The expression of FBXO5 (effect size = 5.77) and TP53 (effect size = 3.66) were evaluated using western blotting. (B) The hSCAPs were transfected with oeNC, oeFBXO5, or oeFBXO5+oeTP53 and subsequently cultured in osteogenic induction medium for 14 days. Cell viability was assessed through the CCK-8 assay (effect size = 2.34). (C) Proliferation of cells was determined by the EdU assay (effect size = 2.49). Scale bar, 100 µm. Data were presented as mean ± SD. ns, no significance, Exact P values were indicated in the figure, P < .05 was considered statistically significant (Ordinary one-way ANOVA with Tukey's multiple comparisons test).

Article Snippet: After blocking with 5% skim milk, the membranes were incubated overnight at 4°C with anti-FBXO5 antibody (1:1000, ab187144, Abcam), anti-TP53 antibody (1:3000, 10442-1-AP, Proteintech, Wuhan, China), anti-runt-related transcription factor 2 (RUNX2) antibody (1:2000, 20700-1-AP, Proteintech), anti-DLX5 antibody (1:1000, 10592-1-AP, Proteintech), anti-osterix (OSX) antibody (1:1000, 28694-1-AP, Proteintech), anti-osteocalcin (OCN) antibody (1:1000, ab133612, Abcam), or anti-GAPDH antibody (1:1000, ab9485, Abcam).

Techniques: Expressing, Transfection, Incubation, Western Blot, Cell Culture, CCK-8 Assay, EdU Assay

Journal: International Dental Journal

Article Title: FBXO5 alleviates apical periodontitis by facilitating TP53 protein degradation

doi: 10.1016/j.identj.2025.103948

Figure Lengend Snippet: FBXO5 promotes osteogenic differentiation of hSCAPs by negatively regulating TP53. The hSCAPs were transfected with oeNC, oeFBXO5, or oeFBXO5+oeTP53 and then cultured in osteogenic induction medium for 14 days. (A-B) ALP staining and enzyme activity assay were conducted to assess ALP activity (effect size = 4.90 and 2.84, respectively). Scale bar, 100 µm. (C) ARS staining was performed to detect mineralisation, and the stained area (%) was quantified using ImageJ software (effect size = 3.51). Scale bar, 100 µm. (D) Western blotting was used to examine the protein expression levels of RUNX2 (effect size = 4.85), DLX5 (effect size = 3.03), OSX (effect size = 8.75), and OCN (effect size = 10.58). Data were presented as mean ± SD. Exact P values were indicated in the figure, P < .05 was considered statistically significant (Ordinary one-way ANOVA with Tukey's multiple comparisons test).

Article Snippet: After blocking with 5% skim milk, the membranes were incubated overnight at 4°C with anti-FBXO5 antibody (1:1000, ab187144, Abcam), anti-TP53 antibody (1:3000, 10442-1-AP, Proteintech, Wuhan, China), anti-runt-related transcription factor 2 (RUNX2) antibody (1:2000, 20700-1-AP, Proteintech), anti-DLX5 antibody (1:1000, 10592-1-AP, Proteintech), anti-osterix (OSX) antibody (1:1000, 28694-1-AP, Proteintech), anti-osteocalcin (OCN) antibody (1:1000, ab133612, Abcam), or anti-GAPDH antibody (1:1000, ab9485, Abcam).

Techniques: Transfection, Cell Culture, Staining, Enzyme Activity Assay, Activity Assay, Software, Western Blot, Expressing