Journal: Antioxidants

Article Title: Saposhnikovia divaricata Inhibits Inflammation, Oxidative Stress, and Ferroptosis to Alleviate DSS-Induced Ulcerative Colitis

doi: 10.3390/antiox15020258

Figure Lengend Snippet: Network pharmacology analysis identifies p53 as a core ferroptosis-related target of FF in UC. ( A ) Venn diagram illustrating the intersection of FF compound targets with ferroptosis- and UC-related targets. ( B ) Protein–protein interaction (PPI) network of the common targets. Node size and color intensity represent the degree of connectivity, with TP53 (p53) identified as the core target. ( C ) Compound-target-pathway network diagram. The inner pink nodes represent the 38 intersecting targets linking FF, UC, and ferroptosis. ( D ) Gene Ontology (GO) enrichment analysis of the common targets, categorized into Biological Process (BP, red), Cellular Component (CC, green), and Molecular Function (MF, blue). ( E ) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis.

Article Snippet: The p53 antibody was obtained from Boster Biological Technology Co., Ltd. (Pleasanton, CA, USA, Item #BM0101).

Techniques:

Journal: Antioxidants

Article Title: Saposhnikovia divaricata Inhibits Inflammation, Oxidative Stress, and Ferroptosis to Alleviate DSS-Induced Ulcerative Colitis

doi: 10.3390/antiox15020258

Figure Lengend Snippet: FF modulates the expression of ferroptosis-related proteins in colon tissue via the p53 pathway. ( A ) Representative immunohistochemical (IHC) images of p53, SLC7A11, and GPX4 expression in colon sections (scale bar = 50 μm). ( B – D ) Quantitative analysis of the relative protein expression levels of p53 (B), SLC7A11 (C), and GPX4 (D). Data are presented as the mean ± SD ( n = 3 independent experiments). ### p < 0.001 versus the control (CON) group; * p < 0.05, ** p < 0.01, *** p < 0.001 versus the DSS model group.

Article Snippet: The p53 antibody was obtained from Boster Biological Technology Co., Ltd. (Pleasanton, CA, USA, Item #BM0101).

Techniques: Expressing, Immunohistochemical staining, Control

Journal: iScience

Article Title: Tumor-suppressive role of the musculoaponeurotic fibrosarcoma gene in colorectal cancer

doi: 10.1016/j.isci.2023.106478

Figure Lengend Snippet: Inverse relationship between c-MAF and p53 protein expression

Article Snippet: . pCMV6-c-MAF plasmid DNA was purchased from OriGene (Rockville, MD, USA). pCMV6-empty vector was used as a control. pCMV-Neo-Bam p53 R175H (R175H), pCMV-Neo-Bam p53 R248W (R248W), and pCMV-Neo-Bam (Empty) were purchased from Addgene ( a gift from Bert Vogelstein, addgene, #16436; http://n2t.net/addgene:16436 ; RRID:Addgene_16436, #16437; http://n2t.net/addgene:16437 ; RRID:Addgene_16437, #16440; http://n2t.net/addgene:16440 ; RRID:Addgene_16440, respectively).

Techniques: Expressing

Journal: iScience

Article Title: Tumor-suppressive role of the musculoaponeurotic fibrosarcoma gene in colorectal cancer

doi: 10.1016/j.isci.2023.106478

Figure Lengend Snippet: Key resources table

Article Snippet: . pCMV6-c-MAF plasmid DNA was purchased from OriGene (Rockville, MD, USA). pCMV6-empty vector was used as a control. pCMV-Neo-Bam p53 R175H (R175H), pCMV-Neo-Bam p53 R248W (R248W), and pCMV-Neo-Bam (Empty) were purchased from Addgene ( a gift from Bert Vogelstein, addgene, #16436; http://n2t.net/addgene:16436 ; RRID:Addgene_16436, #16437; http://n2t.net/addgene:16437 ; RRID:Addgene_16437, #16440; http://n2t.net/addgene:16440 ; RRID:Addgene_16440, respectively).

Techniques: Recombinant, Transfection, DNA Extraction, CCK-8 Assay, SYBR Green Assay, Plasmid Preparation, Amplification, Extraction, Bradford Protein Assay, Microarray, RNA Sequencing, Negative Control, Expressing, Sequencing, Software, CRISPR

Journal: Cell Communication and Signaling : CCS

Article Title: The mTORC2 component SIN1 post-transcriptionally regulates TYMS levels and modulates P53 activity in response to 5-FU chemotherapy

doi: 10.1186/s12964-025-02640-y

Figure Lengend Snippet: SIN1 controls AKT activity and TYMS levels following genotoxic stress. A Lysates from MDA-MB-231 cells stably expressing Sh control or small hairpin against SIN1, treated or not with 50 µM 5-FuDR or 100 µM 5-FU for 24 h, were stained with the indicated antibodies. n = 3 separate biological replicates. B-D Bar graphs showing the results of densitometric analysis of the protein levels under different conditions obtained in A. The data are presented as the means ± SEMs. ANOVA, multiple comparisons: Dunnett test. p-AKT1 (F (5, 12) = 16.02, P < 0.001), p-P70SK (F(5,12) = 2.86, p = 0.06), P53 (F (5, 12) = 3.498, P < 0.05, P21 (F (5, 12) = 26.09, P < 0.0001), TYMS (F (5, 12) = 19.39, P < 0.001). E Bar graphs showing the percentage of Sh Ctrl and Sh SIN1 MDA-MB-231 cells at SubG1 after 72 h of treatment with FuDR. Data are presented as the means ± SEMs. ANOVA (F (3, 8) = 45.1, P < 0.0001). F Lysates of MCF7 cells stably expressing Sh control or small hairpin against SIN1 treated or not with 50 µM 5-FuDR for 24 h, then stained with the indicated antibodies. G-I Bar graphs showing the results of densitometric analysis of the protein levels under different conditions obtained in F. The data are presented as the means ± SEMs. ANOVA, multiple comparisons: Dunnett test. p-AKT1 (F (3, 8) = 483.1, P < 0.0001), p-P70SK (F (3, 8) = 11.58, P < 0.01), P53 (F (3, 8) = 616.3, P < 0.0001), P21 (F (3, 8) = 251.3, P < 0.0001), TYMS (F (3, 8) = 97.82, P < 0.0001). J Bar graphs showing the percentage of MCF7 Sh Ctrl and Sh SIN1 cells at different phase of the cell cycle after 72 h of treatment with FuDR. Data are presented as the means ± SEMs. ANOVA (F (3, 18) = 11.89, ** P < 0.001, * P < 0.05)

Article Snippet: Empty vector c-Flag pcDNA3 was a gift from Stephen Smale (Addgene plasmid # 20011; http://n2t.net/addgene:20011 ; RRID: Addgene_20011). pcDNA3 flag p53 was a gift from Thomas Roberts (Addgene plasmid # 10838; http://n2t.net/addgene:10838 ; RRID:Addgene 10838). p53 (dominant negative R175H mutant)-pcw107-V5 was a gift from David Sabatini & Kris Wood (Addgene plasmid # 64638; http://n2t.net/addgene:64638 ; RRID: Addgene_64638).

Techniques: Activity Assay, Stable Transfection, Expressing, Control, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: The mTORC2 component SIN1 post-transcriptionally regulates TYMS levels and modulates P53 activity in response to 5-FU chemotherapy

doi: 10.1186/s12964-025-02640-y

Figure Lengend Snippet: P53 controls AKT phosphorylation and TYMS levels following genotoxic stress. A , C Lysates from MDA-MB-231 and MCF7 cells stably expressing Sh control or small hairpin against p53, treated or not with 50 µM during 24 h, were stained with the indicated antibodies. B-D Bar graphs showing the results of densitometric analysis of AKT phosphorylation and TYMS protein levels under different conditions obtained in A and C. The data are presented as the means ± SEMs. B ANOVA, multiple comparisons: Dunnett test. p-AKT1 (F (3, 8) = 5.51, P < 0.05), TYMS (F (3, 8) = 240.97, P < 0.0001). D ANOVA, multiple comparisons: Dunnett test. p-AKT1 (F (3, 8) = 291.1, P < 0.001), TYMS (F (3, 8) = 25.51, P < 0.0001)

Article Snippet: Empty vector c-Flag pcDNA3 was a gift from Stephen Smale (Addgene plasmid # 20011; http://n2t.net/addgene:20011 ; RRID: Addgene_20011). pcDNA3 flag p53 was a gift from Thomas Roberts (Addgene plasmid # 10838; http://n2t.net/addgene:10838 ; RRID:Addgene 10838). p53 (dominant negative R175H mutant)-pcw107-V5 was a gift from David Sabatini & Kris Wood (Addgene plasmid # 64638; http://n2t.net/addgene:64638 ; RRID: Addgene_64638).

Techniques: Phospho-proteomics, Stable Transfection, Expressing, Control, Staining

Journal: Cell Communication and Signaling : CCS

Article Title: The mTORC2 component SIN1 post-transcriptionally regulates TYMS levels and modulates P53 activity in response to 5-FU chemotherapy

doi: 10.1186/s12964-025-02640-y

Figure Lengend Snippet: SIN1 controls P53 transcriptional activity following genotoxic stress. A-D Bar graphs showing mRNA levels of TYMS and P53 target genes under the indicated conditions in MCF7 cells. The data are presented as the means ± SEMs. ANOVA, multiple comparisons: Dunnett test. TYMS F (5, 12) = 17.18, P < 0.0001. CDKN1A F (5, 12) = 274.7, P < 0.0001. GAD45A F (5, 12) = 205.0, P < 0.0001. SESN2 F (5, 11) = 12.43, P = 0.0003. E–G Bar graphs showing mRNA levels of P53 target genes under the indicated conditions in MDA-MB-231 cells. The data are presented as the means ± SEMs. ANOVA, multiple comparisons: Dunnett test. CDKN1A F(3,8) = 6.94, P = 0.012. GAD45A F (3, 8) = 22.4, P < 0.001. SESN2 F (3, 8) = 2.22, P = 0.16

Article Snippet: Empty vector c-Flag pcDNA3 was a gift from Stephen Smale (Addgene plasmid # 20011; http://n2t.net/addgene:20011 ; RRID: Addgene_20011). pcDNA3 flag p53 was a gift from Thomas Roberts (Addgene plasmid # 10838; http://n2t.net/addgene:10838 ; RRID:Addgene 10838). p53 (dominant negative R175H mutant)-pcw107-V5 was a gift from David Sabatini & Kris Wood (Addgene plasmid # 64638; http://n2t.net/addgene:64638 ; RRID: Addgene_64638).

Techniques: Activity Assay

Journal: Cell Communication and Signaling : CCS

Article Title: The mTORC2 component SIN1 post-transcriptionally regulates TYMS levels and modulates P53 activity in response to 5-FU chemotherapy

doi: 10.1186/s12964-025-02640-y

Figure Lengend Snippet: Increase in TYMS levels alters P53 activity upon genotoxic stress. A Lysates from MCF7 cells stably expressing Sh control or small hairpin against SIN1 transiently transfected with TYMS expressing plasmid, treated or not with 50 µM during 24 h, were stained with the indicated antibodies. n = 3 separate biological replicates. B Bar graphs showing the results of densitometric analysis of P53 protein levels under different conditions obtained in A. The data are presented as the means ± SEMs. ANOVA, multiple comparisons: Dunnett test. P53 F(7,17) = 44.62, P < 0.0001. C Bar graph displaying the proportion of cells at different stages in the cell cycle analyzed by flow cytometry. MCF7 sh control and sh SIN1 transfected as in A, then treated with or without 50 µM during 72 h. ANOVA, multiple comparisons: Dunnett test. G1 phase F (F (7, 24) = 25.65, P < 0.0001. See supplementary Table 2 for detailed statistics. D Lysates from MCF7 cells treated or not with FuDR for the indicated durations (hours). N = 3 biological replicates per condition. E Bar graph displaying the amount of SIN1 isoforms under the condition obtained in D. The data are presented as the means ± SEMs. ANOVA, multiple comparisons: Dunnett test. Long F(3,8) = 39.67, P < 0.001. Short Long F (3,8) = 26.06, P < 0.001

Article Snippet: Empty vector c-Flag pcDNA3 was a gift from Stephen Smale (Addgene plasmid # 20011; http://n2t.net/addgene:20011 ; RRID: Addgene_20011). pcDNA3 flag p53 was a gift from Thomas Roberts (Addgene plasmid # 10838; http://n2t.net/addgene:10838 ; RRID:Addgene 10838). p53 (dominant negative R175H mutant)-pcw107-V5 was a gift from David Sabatini & Kris Wood (Addgene plasmid # 64638; http://n2t.net/addgene:64638 ; RRID: Addgene_64638).

Techniques: Activity Assay, Stable Transfection, Expressing, Control, Transfection, Plasmid Preparation, Staining, Flow Cytometry

Journal: Journal of Biological Chemistry

Article Title: Mutant p53 Disrupts Role of ShcA Protein in Balancing Smad Protein-dependent and -independent Signaling Activity of Transforming Growth Factor-β (TGF-β)

doi: 10.1074/jbc.m111.265397

Figure Lengend Snippet: FIGURE 1. Mutant p53 inhibits cell migration and ERK signaling in pros- tate cancer cell lines. A, immunoblotting analysis of the indicated gene expression in DU145 cells, which were transfected with control or p53 siRNA for 72 h. B, cell migration was assessed in DU145 cells 48 h after the transfec- tion. Cells were plated at 40,000 cells/insert and incubated for 18 h as described under “Experimental Procedures.” Data represent mean S.E. from three inserts. *, p 0.05. C, immunoblotting analysis of the indicated gene expression in PC-3 cells with stable introduction of p53 R175H expres- sion plasmid (PC-3/mp53) or empty vector. D, cell migration was assessed in PC-3/mp53 and control cells plated at 70,000 cells/insert and incubated for 18 h. Data represent mean S.E. from three inserts. *, p 0.05. T-ERK, total ERK. Error bars represent S.E.

Article Snippet: Plasmids and Transfection—p52/46ShcA plasmid was purchased from Origene. p53 R175H-pCMV-Neo-Bam plasmid was provided by Dr. Harikrishna Nakshatri. p53 R273H-pCDNA3 plasmid was provided by Dr. Zhi-Min Yuan.

Techniques: Mutagenesis, Migration, Western Blot, Gene Expression, Transfection, Control, Incubation, Plasmid Preparation

Journal: Journal of Biological Chemistry

Article Title: Mutant p53 Disrupts Role of ShcA Protein in Balancing Smad Protein-dependent and -independent Signaling Activity of Transforming Growth Factor-β (TGF-β)

doi: 10.1074/jbc.m111.265397

Figure Lengend Snippet: FIGURE 2. Mutant p53 inhibits oncogenic role of TGF- and enhances TGF-/Smad signaling. A, cell migration was assayed with PC-3/mp53 and PC-3 control cells plated at 70,000 cells/insert with or without TGF-1 (2 ng/ml) for 18 h. Data represent mean S.E. from three inserts. **, p 0.01; ***, p 0.0001. B, soft agar colony formation ability was assessed in PC-3/mp53 and PC-3 control cells under the indicated TGF-1 or HTS466284 (HTS) treatment for 12 days. Data represent mean S.E. from three wells. *, p 0.05; **, p 0.01. C, representative pictures of soft agar colonies of PC-3/mp53 and PC-3 control cells were taken after 12-day treatment with 2 ng/ml TGF-1 or 0.1 M HTS466284. D, effect of TGF-1 on cell proliferation was measured with the MTT assay at day 6 after incubation with the indicated dose of TGF-1. Data represent mean S.E. from five wells. A two-tailed t test was performed to compare the mean of relative cell number between PC-3/mp53 and PC-3 control cells for all treatments. *, p 0.05. E, immunoblotting analysis for the indicated gene expression was performed in PC-3/mp53 and PC-3 control cells under TGF-1 treatment for 40 min. F, SBE-luciferase assay was performed using PC-3/mp53 and PC-3 control cells after TGF-1 treatment as described under “Experimental Procedures.” Data represent mean S.E. from three independent transfections of relative luciferase units normalized to -galactosidase activity. *, p 0.05; **, p 0.01. T-Smad2/3, total Smad2/3. Error bars represent S.E.

Article Snippet: Plasmids and Transfection—p52/46ShcA plasmid was purchased from Origene. p53 R175H-pCMV-Neo-Bam plasmid was provided by Dr. Harikrishna Nakshatri. p53 R273H-pCDNA3 plasmid was provided by Dr. Zhi-Min Yuan.

Techniques: Mutagenesis, Migration, Control, MTT Assay, Incubation, Two Tailed Test, Western Blot, Gene Expression, Luciferase, Transfection, Activity Assay

Journal: Journal of Biological Chemistry

Article Title: Mutant p53 Disrupts Role of ShcA Protein in Balancing Smad Protein-dependent and -independent Signaling Activity of Transforming Growth Factor-β (TGF-β)

doi: 10.1074/jbc.m111.265397

Figure Lengend Snippet: FIGURE3.ShcA-mediatedERKsignalingisdown-regulatedinpresenceof mutated p53. A, immunoblotting analysis for the indicated gene expression was performed in PC-3/mp53 and PC-3 control cells after TGF-1 treatment for 40 min. B, immunoblotting analysis for p-ERK and p53 was performed in PC-3 cells 72 h after transfection of p53 R175H expression plasmid, human WTp53 expression plasmid, or the empty vector. C, immunoblotting analysis for the indicated gene expression was performed in PC-3/mp53 cells 72 h after transfection of p53 siRNA 1, p53 siRNA 2, or control siRNA. D, immuno- blotting analysis for the indicated gene expression was assessed in PC-3 cells 72 h after transfection of a p53 R273H expression plasmid or empty vector followed by TGF-1 treatment for 40 min. E, immunoblotting analysis for the indicated gene expression was assessed in PC-3 cells 72 h after transfection of a human WTp53 expression plasmid or empty vector followed by TGF-1 treatment for 40 min. F, immunoblotting analysis for the indicated gene expression was conducted in PC-3/mp53 and PC-3 control cells and PC-3 cells transfected with p53 R273H or WTp53 expression plasmid for 72 h followed by TGF-1 treatment for 40 min. T-ERK, total ERK; T-Smad2/3, total Smad2/3; con; control.

Article Snippet: Plasmids and Transfection—p52/46ShcA plasmid was purchased from Origene. p53 R175H-pCMV-Neo-Bam plasmid was provided by Dr. Harikrishna Nakshatri. p53 R273H-pCDNA3 plasmid was provided by Dr. Zhi-Min Yuan.

Techniques: Western Blot, Gene Expression, Control, Transfection, Expressing, Plasmid Preparation

Journal: Journal of Biological Chemistry

Article Title: Mutant p53 Disrupts Role of ShcA Protein in Balancing Smad Protein-dependent and -independent Signaling Activity of Transforming Growth Factor-β (TGF-β)

doi: 10.1074/jbc.m111.265397

Figure Lengend Snippet: FIGURE 7. Mutant p53 correlates with higher level of Smad-dependent signaling and lower level of ShcA/ERK signaling in human cancer cell lines and genetically altered mouse models. A, immunoblotting analysis for the indicated gene expression was performed in BT-20, BT474, 22Rv1, and DU145 cells 72 h after transfection of p53 siRNA 1 or control siRNA. B, immu- noblotting analysis for the indicated gene expression was performed in the tissuelysatesfrommammaryglandofp53R172H/R172Hpun/un(p53R172H;n2) and p53R172P/R172P pun/un (p53 R172P; n 1) mice and the littermate p53/ pun/un (WTp53; n 5). The scatter plot figures under the immunoblots are the total ERK-normalized p-ERK levels and the total Smad2/3-normalized p-Smad3 levels quantified with ImageJ software. C, immunoblotting analysis for the indicated gene expression was performed in the tissue lysates from mammary tumors of MMTV-Wnt1-WTp53/ (WTp53; n 8), MMTV-Wnt1- p53R172H/R172H (p53 R172H/H; n 4), and MMTV-Wnt1-p53R172H/O (p53 R172H/O; n 4) mice. The scatter plot figures under the immunoblots are the total p52ShcA-normalized p-p52ShcA levels, total ERK-normalized p-ERK lev- els, and the total Smad2/3-normalized p-Smad3 levels for each sample quan- tified with ImageJ software. *, p 0.05. T-ERK, total ERK; T-Smad2/3, total Smad2/3; T-p52ShcA, total p52ShcA.

Article Snippet: Plasmids and Transfection—p52/46ShcA plasmid was purchased from Origene. p53 R175H-pCMV-Neo-Bam plasmid was provided by Dr. Harikrishna Nakshatri. p53 R273H-pCDNA3 plasmid was provided by Dr. Zhi-Min Yuan.

Techniques: Mutagenesis, Western Blot, Gene Expression, Transfection, Control, Software