Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: Glucose and oxygen deprivation can induce endothelial cell senescence. (A) Western blot analysis was performed to quantify the changes in protein levels of Ki-67, Lamin B, FOXO4, P53, P21, P16, and γ-H2AX in HUVECs following OGD treatment, with n ≥ 4 per group. (B) Representative immunofluorescence staining was conducted to analyze the content and localization changes of Ki-67, P21, γ-H2AX, and P53 different groups of HUVECs, with n = 5 per group. (C) Subcellular localization of FOXO4 in different groups of HUVECs was analyzed through representative immunofluorescence staining (FOXO4 in red, DAPI in blue), with n = 5 per group, scale bar = 40 μm. (D) Representative SA-β-GAL staining and quantification were performed to evaluate the number of positive cells in HUVECs after OGD treatment, with n = 5 per group, scale bar = 100 μm. (E) Tube formation assays analyzed relative tube lengths in different groups following OGD treatment, with n = 5 per group, scale bar = 400 μm. (F) Scratch assays were conducted to assess endothelial cell migration coverage at 0, 12, 24, and 36 h across different groups, with n = 5 per group, scale bar = 400 μm. Data are represented as mean ± SEM. Statistical analysis assessed by 2-way RM ANOVA (G) Representative DHE staining and quantification evaluated the number of DHE-positive cells in different groups, with n = 5 per group, scale bar = 100 μm. (H) RT-qPCR analysis was performed in HUVECs following OGD treatment to measure the mRNA levels of IL-6 , IL-8 , IL-1β , and TNF-α , with n = 3 per group. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001.

Article Snippet: The primary antibodies used are as follows: P53 (Proteintech, 60283-2-Ig), pSer46-P53 (CST, 2521), FOXO4 (Proteintech, 21535-1-AP), Ki-67 (D3B5) Rabbit mAb (Alexa Fluor® 647 Conjugate) (CST, 12075), P21 (CST, 14074), γ-H2AX (Abcam, ab2254).

Techniques: Western Blot, Immunofluorescence, Staining, Migration, Quantitative RT-PCR, Two Tailed Test

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: FOXO4-DRI can block the interaction between P53 and FOXO4. (A) The FOXO4-P53 interaction was analyzed through co-immunoprecipitation in OGD-treated HUVECs, in different groups, with n = 3 per group. (B) Western blot analysis quantified changes in protein levels of pSer46-P53 and P53 in different groups, with n = 5 per group. (C) Western blot analysis quantified changes in protein levels of pSer46-P53 and P53 in different groups, with n = 5 per group. (D) Western blot analysis was performed to examine the distribution of pSer46-P53 and P53 in the cytoplasm and nucleus of HUVECs in different groups. With n = 5 per group. (E) Subcellular localization of pSer46-P53 in different groups of HUVECs was analyzed using representative immunofluorescence staining (pSer46-P53 in green, DAPI in blue), with n = 5 per group, scale bar = 40 μm. (F) Immunofluorescence detection and analysis of subcellular localization of pSer46-P53 and FOXO4 in aortic sections of naturally aged mice (pSer46-P53 in green, FOXO4 in red, DAPI in blue), with n = 5 per group, scale bar = 20 μm. (G) Immunofluorescence detection and analysis of subcellular localization of pSer46-P53 and FOXO4 in aortic sections of progeroid mice (pSer46-P53 in green, FOXO4 in red, DAPI in blue), with n = 5 per group, scale bar = 20 μm. Data are presented as mean ± SEM and analyzed using a two-tailed Student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The primary antibodies used are as follows: P53 (Proteintech, 60283-2-Ig), pSer46-P53 (CST, 2521), FOXO4 (Proteintech, 21535-1-AP), Ki-67 (D3B5) Rabbit mAb (Alexa Fluor® 647 Conjugate) (CST, 12075), P21 (CST, 14074), γ-H2AX (Abcam, ab2254).

Techniques: Blocking Assay, Immunoprecipitation, Western Blot, Immunofluorescence, Staining, Two Tailed Test

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: FOXO4-DRI regulates endothelial cell senescence via the P53 signaling pathway

doi: 10.3389/fbioe.2025.1729166

Figure Lengend Snippet: Schematic illustration of FOXO4-DRI in alleviating endothelial cell senescence aging extends cell survival. This study demonstrates that FOXO4-DRI mitigates senescence by inhibiting the FOXO4-P53 interaction, promoting P53 phosphorylation, and inducing apoptosis in senescent cells, thereby alleviating aging-related symptoms. Content produced with BioRender; license VN28TABHO4.

Article Snippet: The primary antibodies used are as follows: P53 (Proteintech, 60283-2-Ig), pSer46-P53 (CST, 2521), FOXO4 (Proteintech, 21535-1-AP), Ki-67 (D3B5) Rabbit mAb (Alexa Fluor® 647 Conjugate) (CST, 12075), P21 (CST, 14074), γ-H2AX (Abcam, ab2254).

Techniques: Phospho-proteomics, Produced

Journal: Iranian Journal of Pharmaceutical Research : IJPR

Article Title: Geranyl Acetate Attenuates Para-phenylenediamine-induced Cytotoxicity, DNA Damage, Apoptosis, and Inflammation in HaCaT Keratinocytes

doi: 10.5812/ijpr-164379

Figure Lengend Snippet: Geranyl acetate (GA) mitigates para-phenylenediamine (PPD)-induced DNA damage response (DDR) signaling in HaCaT cells: A – D, HaCaT cells were seeded in 100 mm dishes (2 × 10 5 cells/dish) and incubated for 24 hours, followed by treatment with GA (0 - 500 μM), in the presence or absence of PPD (250 μM) for 48 hours. Protein expression levels of the DDR-related proteins, including ataxia telangiectasia and Rad3 related protein (ATR), p-ATR, p53, p-p53, p38, p-p38, c-Jun N-terminal kinases (JNK), p-JNK, extracellular signal-regulated kinases (ERK), and p-ERK, were analyzed by Western blotting. The β-Actin was used as a loading control. Protein band intensities were quantified using ImageJ software (version 1.53t). Data are presented as mean ± standard deviation (SD, n = 3). Statistical significance was determined by one-way analysis of variance (ANOVA) followed by Tukey’s post-hoc test (## P < 0.01 and ### P < 0.001 compared with the solvent-treated vehicle control group. * P < 0.05, ** P < 0.01, and *** P < 0.001 compared with the PPD-treated negative control group).

Article Snippet: p-p53 (Ser46) , Rabbit , 1:1000 , CST (#2521).

Techniques: Incubation, Expressing, Western Blot, Control, Software, Standard Deviation, Solvent, Negative Control

Journal: Iranian Journal of Pharmaceutical Research : IJPR

Article Title: Geranyl Acetate Attenuates Para-phenylenediamine-induced Cytotoxicity, DNA Damage, Apoptosis, and Inflammation in HaCaT Keratinocytes

doi: 10.5812/ijpr-164379

Figure Lengend Snippet: Geranyl acetate (GA) improves para-phenylenediamine (PPD)-induced apoptosis and inflammation signaling in HaCaT cells: A and B, HaCaT cells were seeded in 100 mm dishes (2 × 10 5 cells/dish) and incubated for 24 hours, followed by treatment with GA (0 - 500 μM), in the presence or absence of PPD (250 μM) for 48 hours. Protein expression levels of the (A) apoptosis-related proteins [BAX, p53 upregulated modulator of apoptosis (PUMA), cytochrome c, and cleaved PARP] and (B) inflammation-related proteins [signal transducer and activator of transcription 3 (STAT3), p-STAT3, p65, p-p65, NF-kappa-B inhibitor alpha (IκB-α), and p-IκB-α] were analyzed by Western blotting. The β-Actin was used as a loading control. Protein band intensities were quantified using ImageJ software (version 1.53t). Data are presented as mean ± standard deviation (SD, n = 3). Statistical significance was determined by one-way analysis of variance (ANOVA) followed by Tukey’s post-hoc test (### P < 0.001 compared with the solvent-treated vehicle control group. * P < 0.05, ** P < 0.01, and *** P < 0.001 compared with the PPD-treated negative control group).

Article Snippet: p-p53 (Ser46) , Rabbit , 1:1000 , CST (#2521).

Techniques: Incubation, Expressing, Western Blot, Control, Software, Standard Deviation, Solvent, Negative Control

Journal: Journal of Inflammation Research

Article Title: Gegen Qinlian Decoction Ameliorated DSS-Induced Colitis by Attenuating Inflammation, Restoring Intestinal Mucosal Barrier and Modulating Gut Microbiota

doi: 10.2147/JIR.S505423

Figure Lengend Snippet: Western blot analysis of signaling pathways in colon tissues from DSS-induced colitis mice treated with GQD. ( A ) Representative blots for the levels of key proteins (p38, JNK, and ERK) on MAPK signaling pathway. ( B – D ) Quantification of the phosphorylation ratios for p38, JNK, and ERK. ( E ) Representative blots for the verification of hub proteins from network pharmacology. ( F – H ) Quantification of the phosphorylation ratios for STAT3, Akt, and p53. The significant difference is represented by different lowercase letters (a, b, and c) between treatments at p < 0.05.

Article Snippet: The rabbit monoclonal antibody of p-AKT (9271L), AKT (9272T), p-JNK (4668T), JNK (925ZS), p-ERK (4370), ERK (4695), p53 (9282), p-p53 (2521) was purchased from Cell Signaling Technology Co. Ltd (Danvers, Massachusetts, USA).

Techniques: Western Blot, Protein-Protein interactions, Phospho-proteomics