sc 8697 oct4  (Santa Cruz Biotechnology)

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Saxifraga stolonifera inhibits porcine epidemic diarrhea virus infection by disrupting nucleocapsid protein-p53 interaction

doi: 10.3389/fcimb.2025.1615300

Figure Lengend Snippet: S. stolonifera exerts antiviral effects by interfering with the p53-dream signaling pathway in Vero E6 cells. (A) Analysis of the expression levels of p53-DREAM signalling pathway-related proteins in Vero E6 cells. (B) The relative expression levels of p53. (C) p21 (D) p107 (E) p130 (F) p130 phosphorylated (Ser672) (G) p130 phosphorylated (Ser952) (H) E2F4 (I) CyclinA were analyzed and were shown in the graphs. The results are representative of three independent experiments (n=3). Data were represented as mean ± SEM. Student’s t test and one-way ANOVA with multiple comparisons was used for data analysis.

Article Snippet: GAPDH Polyclonal antibody (10494-1-AP), p107 (13354-1-AP), and E2F4 (10923-1-AP) were obtained from Proteintech Group (Wuhan, China).

Techniques: Expressing

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Saxifraga stolonifera inhibits porcine epidemic diarrhea virus infection by disrupting nucleocapsid protein-p53 interaction

doi: 10.3389/fcimb.2025.1615300

Figure Lengend Snippet: S. stolonifera exerts antiviral effects by interfering with the p53-dream signaling pathway in IPEC-J2 cells. (A) Analysis of the expression levels of p53-DREAM signalling pathway-related proteins in IPEC-J2 cells. (B) The relative expression levels of p53. (C) p21 (D) p107 (E) p130 (F) p130 phosphorylated (Ser672) (G) p130 phosphorylated (Ser952) (H) E2F4 (I) CyclinA were analyzed and were shown in the graphs. The results are representative of three independent experiments (n=3). Data were represented as mean ± SEM. Student’s t test and one-way ANOVA with multiple comparisons was used for data analysis.

Article Snippet: GAPDH Polyclonal antibody (10494-1-AP), p107 (13354-1-AP), and E2F4 (10923-1-AP) were obtained from Proteintech Group (Wuhan, China).

Techniques: Expressing

Journal: Experimental and Therapeutic Medicine

Article Title: Aurora kinase A promotes epithelial‑mesenchymal transition by regulating P130 and P107 molecules in thyroid cancer cells

doi: 10.3892/etm.2025.12843

Figure Lengend Snippet: AURKA inhibition can reduce the proliferation by regulating P130 and P107 molecules. (A) Immunofluorescent staining assay was used to detect the expression levels of P130 and P107 in BTH101/si-NC, BTH101/si-AURKA, BCPAP/si-NC and BCPAP/si-AURKA cells. (B) Western blot analysis was used to detect the expression levels of P107, P130, E2F4 and GAPDH in BTH101/si-NC, BTH101/si-AURKA, BCPAP/si-NC and BCPAP/si-AURKA cells. (C) The protein expression level relative to GAPDH in BTH101/si-NC, BTH101/si-AURKA, BCPAP/si-NC and BCPAP/si-AURKA cells. (D) Western blot analysis was used to detect the expression of AURKA in BTH101/vector, BTH101/AURKA, BCPAP/vector and BCPAP/AURKA cells. (E) The protein expression level relative to GAPDH in BTH101/vector, BTH101/AURKA, BCPAP/vector and BCPAP/AURKA cells. (F) Western blot analysis was used to detect the expression levels of P107, P130, E2F4 and GAPDH in BTH101/vector, BTH101/AURKA, BCPAP/vector and BCPAP/AURKA cells. (G) The protein expression level relative to GAPDH in BTH101/vector, BTH101/AURKA, BCPAP/vector and BCPAP/AURKA cells. ** P<0.01. AURKA, Aurora kinase A; siRNA, small interfering RNA; NC, negative control.

Article Snippet: The slides were then rinsed three times with PBS, blocked with 5% BSA for 1 h at 37˚C, and incubated overnight at 4˚C with anti-AURKA (1:100; cat. no. 66757-1-Ig; Cell Signaling Technology, Inc.), anti-P130 (1:100; cat. no. 13383; Cell Signaling Technology, Inc.) and anti-P107 (1:100; cat. no. 13354-1-AP; Proteintech Group, Inc.).

Techniques: Inhibition, Staining, Expressing, Western Blot, Plasmid Preparation, Small Interfering RNA, Negative Control

Journal: Experimental and Therapeutic Medicine

Article Title: Aurora kinase A promotes epithelial‑mesenchymal transition by regulating P130 and P107 molecules in thyroid cancer cells

doi: 10.3892/etm.2025.12843

Figure Lengend Snippet: AURKA promotes epithelial-mesenchymal transition through the FAK signaling pathway and FAK pathway inhibition impacts proliferation-related proteins. (A) Western blot analysis was used to detect the expression of FAK in BTH101/si-NC, BTH101/si-AURKA, BCPAP/si-NC and BCPAP/si-AURKA cells. (B) Western blot analysis was used to detect the expression of FAK in BTH101/vector, BTH101/AURKA, BCPAP/vector and BCPAP/AURKA cells. (C) The protein expression level relative to GAPDH in BTH101/si-NC, BTH101/si-AURKA, BCPAP/si-NC and BCPAP/si-AURKA cells. (D) The protein expression level relative to GAPDH in BTH101/vector, BTH101/AURKA, BCPAP/vector and BCPAP/AURKA cells. (E) Western blot analysis was used to detect the expression levels of P107, P130, E2F4 and GAPDH in BTH101/si-NC, BTH101/si-AURKA, BCPAP/si-NC and BCPAP/si-AURKA cells. (F) Western blot analysis was used to detect the expression levels of P107, P130, E2F4 and GAPDH in BTH101/vector, BTH101/AURKA, BCPAP/vector and BCPAP/AURKA cells. (G) The protein expression level relative to GAPDH in BTH101/si-NC, BTH101/si-AURKA, BCPAP/si-NC and BCPAP/si-AURKA cells. (H) The protein expression level relative to GAPDH in BTH101/vector, BTH101/AURKA, BCPAP/vector and BCPAP/AURKA cells. ** P<0.01. AURKA, Aurora kinase A; FAK, focal adhesion kinase; siRNA, small interfering RNA; NC, negative control.

Article Snippet: The slides were then rinsed three times with PBS, blocked with 5% BSA for 1 h at 37˚C, and incubated overnight at 4˚C with anti-AURKA (1:100; cat. no. 66757-1-Ig; Cell Signaling Technology, Inc.), anti-P130 (1:100; cat. no. 13383; Cell Signaling Technology, Inc.) and anti-P107 (1:100; cat. no. 13354-1-AP; Proteintech Group, Inc.).

Techniques: Inhibition, Western Blot, Expressing, Plasmid Preparation, Small Interfering RNA, Negative Control

Journal: The EMBO Journal

Article Title: p53 regulates DREAM complex-mediated repression in a p21-independent manner

doi: 10.1038/s44318-025-00402-7

Figure Lengend Snippet: ( A , B ) p53 WT, along with DREAM complex members, bind to BLM promoter. (Left) Input for DNA affinity assay. Nuclear extracts were isolated from asynchronously growing ( A ) HCT116 WT, HCT116 p53−/−, HCT116 p21−/− cells, ( B ) HT1080 WT, HT1080 p53−/−, HT1080 p21−/− cells. Western blots were carried out with the indicated antibodies. (Right) DNA affinity assay was carried out using a biotinylated probe encompassing the 640 bp BLM minimal promoter as the “bait” and the nuclear extracts from ( A ) HCT116 p53 WT, HCT116 p53−/−, HCT116 p21−/− cells, ( B ) HT1080 WT, HT1080 p53−/−, HT1080 p21−/− cells as the “prey”. Immunoblotting was performed using the indicated antibodies. Three biological replicates were carried out, and the same result was obtained. ( C ) Binding of p53 and DREAM complex members occur on the #2 E2F site in the BLM promoter. Same as ( A ), except the DNA affinity assay was carried out only in HCT116 p53 WT cells using a biotinylated probe encompassing either the 640 bp wild type BLM minimal promoter or BLM promoter where #2 E2F binding site is mutated. Three biological replicates were carried out and the same result was obtained. ( D ) Both p53 WT and p53 TAD mutant bind to BLM promoter. Same as ( A ) except the nuclear extracts were made from HCT116 p53−/− cells expressing p53 WT or p53 TAD mutant. Immunoblotting was performed using the indicated antibodies. Three biological replicates were carried out and the same result was obtained. ( E – G ). p107/p130 interact with p53 in cells even in the absence of p21. ( E ) Lysates were made from HCT116 WT, HCT116 p53−/−, HCT116 p21−/− cells. Immunoblotting was carried out with the indicated antibodies. ( F , G ) Immunoprecipitation was carried out with antibodies against either (F) p107 or (G) p130. Immunoprecipitates were probed with the indicated antibodies. Three biological replicates were carried out and the same result was obtained. ( H – J ) p107/p130 interact with both p53 WT and p53 TAD mutant irrespective of their phosphorylation status. ( H ) Lysates were made from inducible stable lines generated in HCT116 p53−/− cells expressing either the vector, Flag-tagged p53 WT or Flag-tagged p53 TAD mutant grown in the presence of Doxycycline. Immunoblotting was carried out with the indicated antibodies ( I , J ) Immunoprecipitation was carried out with ( I ) anti-p107 or ( J ) p130 antibodies, and immunoprecipitates were probed with the indicated antibodies. Three biological replicates were carried out, and the same result was obtained. ( K ) E2F4 and p53 are co-recruited onto the BLM promoter in the absence of p21. Re-ChIP assay was carried out using HCT116 p53 WT, HCT116 p53−/−, HCT116 p21−/− cells using anti-E2F4 as the 1 st antibody and anti-p53 as the 2 nd antibody. Co-recruitment of E2F4 and p53 on the #2 E2F site on the BLM promoter was determined by ChIP-qPCR analysis. E2F4-p53 co-recruitment to the GAPDH promoter was used as a control. Mean ± SD. The data is from three biological replicates. ( L ) E2F4 and p53 are co-recruited onto the BLM promoter in cells expressing either p53 WT or p53 TAD mutant. Similar to ( K ), recruitment of E2F4 and p53 to the #2 E2F site on BLM promoter was determined in the indicated stable lines grown in the presence of Doxycycline. Mean ± SD. The data is from three biological replicates. .

Article Snippet: Rabbit anti-p107 (used for WB) , Proteintech , Cat# 13354-1-AP, RRID: AB_2238024) (WB: 1:1000).

Techniques: Isolation, Western Blot, Binding Assay, Mutagenesis, Expressing, Immunoprecipitation, Phospho-proteomics, Generated, Plasmid Preparation, ChIP-qPCR, Control

Journal: The EMBO Journal

Article Title: p53 regulates DREAM complex-mediated repression in a p21-independent manner

doi: 10.1038/s44318-025-00402-7

Figure Lengend Snippet: ( A , B ) p53 R175H cannot repress DREAM complex targets. ( A ) Lysates were prepared from the stable lines grown in the absence or presence of Doxycycline (Dox). Western analysis was performed with the indicated antibodies. Mean ± SD. Three replicates were carried out and the same result was obtained. ( B ) RNA was isolated from the indicated stable lines grown in the presence of Doxycycline. RT-qPCR of the indicated genes was performed. The data is from three biological replicates. ( C ) E2F4 interacts with cancer-specific hotspots p53 mutants in vitro. Interaction assays were carried out using in vitro-translated E2F4 protein and GST-tagged recombinant p53 variants. The interactions were determined by immunoblotting with anti-E2F4 antibody. Three biological replicates were carried out and the same result was obtained. ( D ) E2F4 interact with p53 R175H in cells. (Left) Input for immunoprecipitation of p130, p107 with p53 WT and p53 R175H. Lysates were prepared from the inducible stable cell lines expressing vector, Flag-tagged p53 WT or Flag-tagged p53 R175H in the presence of Dox. Western blots are carried out with the indicated antibodies. (Right) Immunoprecipitation was carried out with anri-E2F4 antibodies and immunoprecipitates were probed with the indicated antibodies. Three biological replicates were carried out and the same result was obtained. ( E ) p53 hotspot mutants cannot bind to BLM promoter. (Left) Inputs for DNA affinity assay. Nuclear extracts were isolated from HCT116 p53−/− cells expressing either p53 WT, p53 R175H, or p53 R248Q in the presence of Dox. Western blots are carried out with the indicated antibodies. (Right) DNA affinity experiment was performed using a biotinylated probe encompassing the 640 bp wild type BLM minimal promoter. Nuclear extracts were made. Immunoblotting was performed using the indicated antibodies. Three biological replicates were carried out, and the same result was obtained. ( F ) E2F4 does not bind to the #2 E2F site on the BLM promoter in the presence of p53 R175H. Recruitment of E2F4 to the #2 E2F site on BLM promoter was determined by ChIP-qPCR in stable lines expressing either vector, p53 WT or p53 R175H, grown in the presence of Doxycycline. The data is from three biological replicates. Mean ± SD. ( G ) E2F4 and p53 are not co-recruited onto the BLM promoter in cells expressing p53 R175H. Re-ChIP assay was carried out using stable lines expressing either the Vector, p53 WT or p53 R175H, grown in the presence of Doxycycline. Anti-E2F4 was used as the 1 st antibody and anti-p53 as the 2 nd antibody. Co-recruitment of E2F4 and p53 on the #2 E2F site on the BLM promoter was determined by ChIP-qPCR analysis. E2F4-p53 co-recruitment to the GAPDH promoter was used as a control. Mean ± SD. The data is from three biological replicates. .

Article Snippet: Rabbit anti-p107 (used for WB) , Proteintech , Cat# 13354-1-AP, RRID: AB_2238024) (WB: 1:1000).

Techniques: Western Blot, Isolation, Quantitative RT-PCR, In Vitro, Recombinant, Immunoprecipitation, Stable Transfection, Expressing, Plasmid Preparation, ChIP-qPCR, Control

Journal: The EMBO Journal

Article Title: p53 regulates DREAM complex-mediated repression in a p21-independent manner

doi: 10.1038/s44318-025-00402-7

Figure Lengend Snippet: p53 represses the DREAM complex by both “canonical” and “non-canonical” pathways. The expression of either wild type p53 or transactivation-dead p53 in normal cells presents two potential outcomes. In the first scenario, wild type p53 activates the cyclin-dependent kinase inhibitor p21, resulting in the hypo-phosphorylation of p107 and p130. This leads to the formation of the DREAM complex, which represses its targets through the “canonical pathway”. However, both wild type and transactivation-dead p53 can interact with p130/p107 irrespective of their phosphorylation levels, through which p53 gets recruited on the DREAM complex promoters and represses the DREAM complex targets through the “non-canonical pathway”. In cancer cells with bi-allelic loss of p53 or the presence of p53 “hotspot mutants”—different mechanisms come into play. Bi-allelic loss of p53 prevents the activation of p21, thus inhibiting the formation of the repressive DREAM complex. Similarly, p53 “hotspot mutants” physically bind and sequester E2F4, preventing the formation of the repressive DREAM complex. Hence in case of either bi-allelic loss of p53 or the presence of p53 “hotspot mutants,”—transcriptional activation occur of the DREAM complex targets.

Article Snippet: Rabbit anti-p107 (used for WB) , Proteintech , Cat# 13354-1-AP, RRID: AB_2238024) (WB: 1:1000).

Techniques: Expressing, Phospho-proteomics, Activation Assay

Journal: The EMBO Journal

Article Title: p53 regulates DREAM complex-mediated repression in a p21-independent manner

doi: 10.1038/s44318-025-00402-7

Figure Lengend Snippet: Reagents and tools table

Article Snippet: Rabbit anti-p107 (used for WB) , Proteintech , Cat# 13354-1-AP, RRID: AB_2238024) (WB: 1:1000).

Techniques: Reverse Transcription, SYBR Green Assay, Extraction, Bicinchoninic Acid Protein Assay, Plasmid Preparation, Mutagenesis, Recombinant, Cloning, Expressing, Control, Sequencing, Software