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: Aging cell

Article Title: Positive crosstalk between arginase-II and S6K1 in vascular endothelial inflammation and aging.

doi: 10.1111/acel.12001

Figure Lengend Snippet: Fig. 1 Silencing Arg-II in senescent cells inhibits eNOS uncoupling, reverses endothelial senescent phenotypic changes, and suppresses endothelial inflammation. Senescent human umbilical vein endothelial cells (HUVECs) were transduced with rAd ⁄ U6-LacZshRNA as control (con) or rAd ⁄ U6-Arg-IIshRNA to silence Arg-II gene. (A) Immunoblotting shows Arg-II silencing in senescent cells. (B) DHE staining for detection of O 2 and DAF-2DA staining for detection of NO. Quantifications of DHE and DAF-2DA signals are shown below. (C) SA-b-gal staining. Bar graphs show quantifications of SA-b-gal-positive cells. (D) Immunoblotting analysis of senescence markers p53-S15, p53, and p21Cip1

Article Snippet: Antibody against p21Cip1 (OP64) was purchased from Calbiochem (Genève, Switzerland); antibody against phosphor-p53-S15 (#9284s) was from Cell Signalling (Allschwil, Switzerland); Antibodies against S6K1 (#9205s) were from BD Transduction laboratories (Allschwil, Switzerland); antibodies against arginase-II (sc-20151) and p53 (sc-6243) were from SantaCruz (Nunningen, Switzerland); anti-SOD1 (TA302692) was from OriGene Technologies, Inc (Nunningen, Switzerland); Monoclonal antibody against HA-tag (12CA5) was obtained from Dr Brian A. Hemmings (Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland); Alexa Fluor680-conjugated anti-mouse IgG (A21057) and dihydroethidium (DHE) were from Molecular Probes ⁄ Invitrogen (Lucerne, Switzerland); IRDye800-conjugated anti-rabbit IgG (926-32211) were from LI-COR Biosciences (Bad Homburg, Germany); the membrane-permeable 4,5- diaminofluoresceine acetate (DAF-2DA) was from VWR international SA (Dietikon, Switzerland); X-gal was from Promega (Dübendorf, Switzerland); Endothelial cell growth supplement (ECGS) pack was from PromoCell GmbH (Allschwil, Switzerland) and all cell culture media and materials were purchased from Gibco BRL (Lucerne, Switzerland).

Techniques: Transduction, Control, Western Blot, Staining

Journal: Aging cell

Article Title: Positive crosstalk between arginase-II and S6K1 in vascular endothelial inflammation and aging.

doi: 10.1111/acel.12001

Figure Lengend Snippet: Fig. 2 Co-expression of superoxide dismutase-1 (SOD1) in young endothelial cells prevents Arg-II-induced eNOS-uncoupling, endothelial senescence and inflammation. Young endothelial cells were transduced with empty rAd ⁄ CMV vector as control (con) or rAd ⁄ CMV-Arg-II alone or rAd ⁄ CMV-Arg-II plus rAd ⁄ CMV-SOD1. (A) Immunoblotting analysis to confirm overexpression of Arg-II and SOD1. (B) DHE staining for detection of O 2 and DAF-2DA staining for detection of NO and effect of SOD1. Bar graphs show quantifications of DHE and DAF-2DA signals. (C) SA-b-gal staining and effect of SOD1. Bar graphs show quantifications of percentage of SA-b-gal positive cells. (D) Immunoblotting analysis of senescence markers p53-S15, p53, and p21Cip1 levels, and endothelial inflammation markers vascular adhesion molecule-1 (VCAM1) and intercellular adhesion molecule-1 (ICAM1) expression. Tubulin served as loading control. Bar graphs show quantifications of the markers. (E) CFDA-SE fluorescence labeled THP-1 monocyte adhesion to endothelial cells that were transduced with rAd expressing transgenes as indicated. Bar graphs show quantifications of the adhered monocytes. *P < 0.05, **P < 0.01 and ***P < 0.005 vs. control (con); ††<0.01 and †††P < 0.005 vs. Arg-II. Scale bar = 0.2 mm.

Article Snippet: Antibody against p21Cip1 (OP64) was purchased from Calbiochem (Genève, Switzerland); antibody against phosphor-p53-S15 (#9284s) was from Cell Signalling (Allschwil, Switzerland); Antibodies against S6K1 (#9205s) were from BD Transduction laboratories (Allschwil, Switzerland); antibodies against arginase-II (sc-20151) and p53 (sc-6243) were from SantaCruz (Nunningen, Switzerland); anti-SOD1 (TA302692) was from OriGene Technologies, Inc (Nunningen, Switzerland); Monoclonal antibody against HA-tag (12CA5) was obtained from Dr Brian A. Hemmings (Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland); Alexa Fluor680-conjugated anti-mouse IgG (A21057) and dihydroethidium (DHE) were from Molecular Probes ⁄ Invitrogen (Lucerne, Switzerland); IRDye800-conjugated anti-rabbit IgG (926-32211) were from LI-COR Biosciences (Bad Homburg, Germany); the membrane-permeable 4,5- diaminofluoresceine acetate (DAF-2DA) was from VWR international SA (Dietikon, Switzerland); X-gal was from Promega (Dübendorf, Switzerland); Endothelial cell growth supplement (ECGS) pack was from PromoCell GmbH (Allschwil, Switzerland) and all cell culture media and materials were purchased from Gibco BRL (Lucerne, Switzerland).

Techniques: Expressing, Transduction, Plasmid Preparation, Control, Western Blot, Over Expression, Staining, Labeling

Journal: Aging cell

Article Title: Positive crosstalk between arginase-II and S6K1 in vascular endothelial inflammation and aging.

doi: 10.1111/acel.12001

Figure Lengend Snippet: Fig. 5 Silencing Arg-II prevents S6K1-induced eNOS-uncoupling, senescence and inflammation in young endothelial cells. The transduction procedure of young cells was the same as in Fig. 5A. (A) DHE staining for detection of O 2 and DAF-2DA staining for detection of NO and effect of Arg-II silencing. Bar graphs show quantifications of DHE and DAF-2DA signals. (B) SA-b-gal staining and effect of Arg-II silencing. Bar graphs show quantifications of percentage of SA-b-gal positive cells. (C) Immunoblotting analysis of senescence markers p53-S15, p53, and p21Cip1 levels, and endothelial inflammation markers vascular adhesion molecule-1 (VCAM1) and intercellular adhesion molecule-1 (ICAM1). Tubulin served as loading control. Bar graphs show quantifications of the markers. (D) CFDA-SE fluorescence labeled THP-1 monocyte adhesion to endothelial cells that were transduced with rAd expressing transgenes and shRNA as indicated. Bar graphs show quantifications of the adhered monocytes. **P < 0.01, ***P < 0.005 vs. control (con ⁄ LacZ); †P < 0.05, ††P < 0.01, †††P < 0.005 vs. S6K1ca ⁄ LacZ group. Scale bar = 0.2 mm.

Article Snippet: Antibody against p21Cip1 (OP64) was purchased from Calbiochem (Genève, Switzerland); antibody against phosphor-p53-S15 (#9284s) was from Cell Signalling (Allschwil, Switzerland); Antibodies against S6K1 (#9205s) were from BD Transduction laboratories (Allschwil, Switzerland); antibodies against arginase-II (sc-20151) and p53 (sc-6243) were from SantaCruz (Nunningen, Switzerland); anti-SOD1 (TA302692) was from OriGene Technologies, Inc (Nunningen, Switzerland); Monoclonal antibody against HA-tag (12CA5) was obtained from Dr Brian A. Hemmings (Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland); Alexa Fluor680-conjugated anti-mouse IgG (A21057) and dihydroethidium (DHE) were from Molecular Probes ⁄ Invitrogen (Lucerne, Switzerland); IRDye800-conjugated anti-rabbit IgG (926-32211) were from LI-COR Biosciences (Bad Homburg, Germany); the membrane-permeable 4,5- diaminofluoresceine acetate (DAF-2DA) was from VWR international SA (Dietikon, Switzerland); X-gal was from Promega (Dübendorf, Switzerland); Endothelial cell growth supplement (ECGS) pack was from PromoCell GmbH (Allschwil, Switzerland) and all cell culture media and materials were purchased from Gibco BRL (Lucerne, Switzerland).

Techniques: Transduction, Staining, Western Blot, Control, Labeling, Expressing, shRNA

Journal: Aging cell

Article Title: Positive crosstalk between arginase-II and S6K1 in vascular endothelial inflammation and aging.

doi: 10.1111/acel.12001

Figure Lengend Snippet: Fig. 6 Deficiency in Arg-II gene in mice (Arg-II) ⁄ )) protects against vascular inflammation and aging. Aortas of young (2–3 months) and old (23–24 months) wild-type (WT) and Arg-II) ⁄ ) mice were cleaned of perivascular tissues and subjected to en face staining or Immunoblotting analysis. (A) qRT-PCR analysis of Arg-II mRNA levels. (B) Confocal microscopic en face detection of endothelial vascular adhesion molecule-1 (VCAM1) and intercellular adhesion molecule-1 (ICAM1), vWF (the endothelial marker), followed by counterstaining with DAPI. Shown are representative images of each group. (C) Immunoblotting analyses of VCAM1, ICAM1, p21 levels in the aortas and p53-Ser15 and p53 levels in the heart of young and old WT and Arg-II) ⁄ ) mice. (D and E) Quantifications of the above results. Tubulin is taken as loading control. n = 4 mice in each group. **P < 0.01, ***P < 0.001 vs. young WT mice; †P < 0.05, ††P < 0.01, †††P < 0.001 vs. old WT mice. Scale bar = 100 lm.

Article Snippet: Antibody against p21Cip1 (OP64) was purchased from Calbiochem (Genève, Switzerland); antibody against phosphor-p53-S15 (#9284s) was from Cell Signalling (Allschwil, Switzerland); Antibodies against S6K1 (#9205s) were from BD Transduction laboratories (Allschwil, Switzerland); antibodies against arginase-II (sc-20151) and p53 (sc-6243) were from SantaCruz (Nunningen, Switzerland); anti-SOD1 (TA302692) was from OriGene Technologies, Inc (Nunningen, Switzerland); Monoclonal antibody against HA-tag (12CA5) was obtained from Dr Brian A. Hemmings (Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland); Alexa Fluor680-conjugated anti-mouse IgG (A21057) and dihydroethidium (DHE) were from Molecular Probes ⁄ Invitrogen (Lucerne, Switzerland); IRDye800-conjugated anti-rabbit IgG (926-32211) were from LI-COR Biosciences (Bad Homburg, Germany); the membrane-permeable 4,5- diaminofluoresceine acetate (DAF-2DA) was from VWR international SA (Dietikon, Switzerland); X-gal was from Promega (Dübendorf, Switzerland); Endothelial cell growth supplement (ECGS) pack was from PromoCell GmbH (Allschwil, Switzerland) and all cell culture media and materials were purchased from Gibco BRL (Lucerne, Switzerland).

Techniques: Staining, Western Blot, Quantitative RT-PCR, Marker, Control

Journal: Journal of Cancer

Article Title: Mutant Pattern of p53 as a Feasible Predictor of Distant Metastasis Following Curative Gastrectomy for Advanced-stage Gastric Cancer

doi: 10.7150/jca.98563

Figure Lengend Snippet: Clinical characteristics of the patients with p53 staining patterns.

Article Snippet: IHC was performed using primary antibody for p53 (clone DO-7, cat. no. ZM0408, 1:200 dilution; Zhongshan GoldenBridge Biotechnology, Beijing, China) and BOND-III autostainer (Leica Biosystems Nussloch GmbH, Nussloch, Germany).

Techniques: Staining, Mutagenesis

Journal: Journal of Cancer

Article Title: Mutant Pattern of p53 as a Feasible Predictor of Distant Metastasis Following Curative Gastrectomy for Advanced-stage Gastric Cancer

doi: 10.7150/jca.98563

Figure Lengend Snippet: Recurrence patterns involving both p53 mutant and wild-type patterns.

Article Snippet: IHC was performed using primary antibody for p53 (clone DO-7, cat. no. ZM0408, 1:200 dilution; Zhongshan GoldenBridge Biotechnology, Beijing, China) and BOND-III autostainer (Leica Biosystems Nussloch GmbH, Nussloch, Germany).

Techniques: Mutagenesis

Journal: Journal of Cancer

Article Title: Mutant Pattern of p53 as a Feasible Predictor of Distant Metastasis Following Curative Gastrectomy for Advanced-stage Gastric Cancer

doi: 10.7150/jca.98563

Figure Lengend Snippet: Overall survival and recurrence-free survival of patients with both the p53 wild-type and mutant patterns. Kaplan-Meier curves for (A) overall survival and (B) recurrence-free survival over a period of 5 years with the p53 wild-type and p53 mutant pattern. Patients with the p53 mutant pattern had low (A) overall survival and (B) recurrence-free survival rates in all patients. In subgroup analysis, the recurrence-free survival rate was lower in patients with the p53 mutant pattern than in those with the wild-type pattern as regards both (C) pN0 or (D) pN+, and (E) early and (F) advanced-stage gastric cancer.

Article Snippet: IHC was performed using primary antibody for p53 (clone DO-7, cat. no. ZM0408, 1:200 dilution; Zhongshan GoldenBridge Biotechnology, Beijing, China) and BOND-III autostainer (Leica Biosystems Nussloch GmbH, Nussloch, Germany).

Techniques: Mutagenesis

Journal: Journal of Cancer

Article Title: Mutant Pattern of p53 as a Feasible Predictor of Distant Metastasis Following Curative Gastrectomy for Advanced-stage Gastric Cancer

doi: 10.7150/jca.98563

Figure Lengend Snippet: Comparison of the characteristics of patients with early- and advanced-stage gastric cancer in association with the p53 wild-type and p53 mutant pattern.

Article Snippet: IHC was performed using primary antibody for p53 (clone DO-7, cat. no. ZM0408, 1:200 dilution; Zhongshan GoldenBridge Biotechnology, Beijing, China) and BOND-III autostainer (Leica Biosystems Nussloch GmbH, Nussloch, Germany).

Techniques: Comparison, Mutagenesis

Journal: Journal of Cancer

Article Title: Mutant Pattern of p53 as a Feasible Predictor of Distant Metastasis Following Curative Gastrectomy for Advanced-stage Gastric Cancer

doi: 10.7150/jca.98563

Figure Lengend Snippet: Clinical characteristics of the patients with p53 staining patterns before and after matching on the propensity score.

Article Snippet: IHC was performed using primary antibody for p53 (clone DO-7, cat. no. ZM0408, 1:200 dilution; Zhongshan GoldenBridge Biotechnology, Beijing, China) and BOND-III autostainer (Leica Biosystems Nussloch GmbH, Nussloch, Germany).

Techniques: Staining, Mutagenesis

Journal: Journal of Cancer

Article Title: Mutant Pattern of p53 as a Feasible Predictor of Distant Metastasis Following Curative Gastrectomy for Advanced-stage Gastric Cancer

doi: 10.7150/jca.98563

Figure Lengend Snippet: Comparison of patient characteristics between pN0 and pN+ gastric cancer in p53 wild-type and mutant pattern.

Article Snippet: IHC was performed using primary antibody for p53 (clone DO-7, cat. no. ZM0408, 1:200 dilution; Zhongshan GoldenBridge Biotechnology, Beijing, China) and BOND-III autostainer (Leica Biosystems Nussloch GmbH, Nussloch, Germany).

Techniques: Comparison, Mutagenesis

Journal: Journal of cellular and molecular medicine

Article Title: Functional role of RRS1 in breast cancer cell proliferation.

doi: 10.1111/jcmm.13922

Figure Lengend Snippet: FIGURE 5 P53 and p21 are induced by RRS1 knockdown. MCF‐ 7 cells were infected with a retrovirus expressing RRS1 (shRRS1) or with a Ctrl vector (shctrl) for 2 days. Whole‐cell lysates were analysed by Western blot. P53 and p21 expression levels were increased by RRS1 knockdown (*P < 0.05 vs shctrl)

Article Snippet: For western blotting, xenograft tumors and cell lines were lysed, and protein samples were harvested as previously described.29 Equal amounts of protein were resolved by SDS‐PAGE and blotted using antibodies specific to RRS1 (1:1000, Abcam), p53 (1:500, OriGene), RPL11 (1:1000, Abcam) and β‐actin (1:1000, Bioss, Beijing, China).

Techniques: Knockdown, Infection, Expressing, Plasmid Preparation, Western Blot

Journal: Archives of Biological Sciences

Article Title: Quercetin and lithium chloride modulate Wnt signaling in pluripotent embryonal carcinoma NT2/D1 cells

doi: 10.2298/abs1301201m

Figure Lengend Snippet: Fig. 2. LiCl decreases the proliferation rate of NT2/D1 cells in a p53-dependent manner. (A) MTT proliferation assay of NT2/D1 treated with lithium chloride-7, 10 and 20 mM, for 24 h. The results are shown as per- centages of the negative control, untreated NT2/D1 cells. Values are presented as the means ±S.E.M. of at least three independent experiments. Mean values of relative proliferation rates were compared using Student’s t test. Values of p<0, 05 are presented by *. (B) Western blot analysis of p53 protein expression in NT2/ D1 cells treated with 7, 10 and 20 mM lithium chloride for 24 h. The level of GAPDH was used as a control for equal amounts of input proteins.

Article Snippet: Western blots were performed using anti c-myc (9E10) (Santa Cruz Biotechnology), anti-p53 (DO1) (Gene Spin), anti α-tubulin (DM1A) (Calbi- ochem), and anti-GAPDH (AM20337PU-S) (Acris Antibodies, Inc).

Techniques: Proliferation Assay, Negative Control, Western Blot, Expressing, Control

Journal: Cancer Research

Article Title: The Common Germline TP53-R337H Mutation Is Hypomorphic and Confers Incomplete Penetrance and Late Tumor Onset in a Mouse Model

doi: 10.1158/0008-5472.can-20-1750

Figure Lengend Snippet: Figure 1. Generation of germline p53-R334H mouse model. A, Targeting strategy. The p53R334H point mutation in exon 10 is marked by an asterisk. B, Identification of targeted ES cell clones by PCR amplification using an outside probe yielding a 3.6 kb WT band and a 4.9 kb mutant band. C, Genotyping of animals using allele-specific primers results in a 109 bp WT band and a 128 bp mutant p53 band. PCR products were resolved on a 3% LMP agarose gel. D, Sequencing of the p53 missense mutation from genomic DNA. E, Genotype analysis of offspring from p53R334H heterozygous crosses.

Article Snippet: Cell lysates were prepared, and total cellular protein was subjected to immunoprecipitation using rabbit polyclonal anti-p53 antibody (FL393; Santa Cruz Biotechnology) followed by immunoblotting as described above using sheep polyclonal anti-p53 (AP05006PU-N; Acris Antibodies, Inc.), mouse monoclonal anti-Mdm2 (2A10, cat. #ab16895; Abcam), and mouse monoclonal anti–b-actin (cat. #A5441; Sigma).

Techniques: Mutagenesis, Clone Assay, Agarose Gel Electrophoresis, Sequencing

Journal: Cancer Research

Article Title: The Common Germline TP53-R337H Mutation Is Hypomorphic and Confers Incomplete Penetrance and Late Tumor Onset in a Mouse Model

doi: 10.1158/0008-5472.can-20-1750

Figure Lengend Snippet: Figure 2. Mutant p53R334H/R334H mice develop tumors with long latency and incom- plete penetrance. A, Kaplan–Meier tumor-free survival curves. B, Histopa- thology of tumors arising in homozy- gous p53R334H mouse (20 months of age). Osteosarcoma, vertebra, hema- toxylin and eosin (2.5, top left plot), osteosarcoma, vertebra, hematoxylin and eosin (40, top middle plot), oste- osarcoma, p53 IHC (40, top right plot); mammary adenocarcinoma, hematoxy- lin and eosin (40, bottom left plot) mammary adenocarcinoma, p53 IHC (40, bottom right plot). C, Histopa- thology of primary and metastatic sar- coma arising in homozygous p53R334H mouse (21 months of age). Angiosar- coma with inset highlighting microcapil- lary formation with red blood cells (40, left), angiosarcoma CD31 staining (40, middle), and metastatic angiosar- coma attached to blood vessel wall (10, right).

Article Snippet: Cell lysates were prepared, and total cellular protein was subjected to immunoprecipitation using rabbit polyclonal anti-p53 antibody (FL393; Santa Cruz Biotechnology) followed by immunoblotting as described above using sheep polyclonal anti-p53 (AP05006PU-N; Acris Antibodies, Inc.), mouse monoclonal anti-Mdm2 (2A10, cat. #ab16895; Abcam), and mouse monoclonal anti–b-actin (cat. #A5441; Sigma).

Techniques: Mutagenesis, Staining

Journal: Cancer Research

Article Title: The Common Germline TP53-R337H Mutation Is Hypomorphic and Confers Incomplete Penetrance and Late Tumor Onset in a Mouse Model

doi: 10.1158/0008-5472.can-20-1750

Figure Lengend Snippet: Figure 3. Mutant p53-R334H is abnormally expressed and functionally impaired in thymocytes. A, FACS analysis of isolated thymocytes in cell culture either untreated (Control) or 16 hours post IR (5 Gy g-IR). Results are representative of four independent experiments. B, Western blot analysis of isolated thymocytes in cell culture at 0 (untreated), 2, and 5 hours post g-IR (5 Gy). Proteins were detected using the following primary antibodies: p53 (1C12); p21 (F5); Puma (P4743); b-actin (AC-15). C, Western blot analysis of tissues from WT, p53R334H/R334H, p53R172H/R172H, andp53/ mice either untreated (Control) or 6 hours after treatment with 5 Gy whole-body IR. Each sample represents an individual animal. Proteins were detected using the following primary antibodies: p53 (1C12); p21 (F5); b-actin (AC-15). D, IHC analysis of p53 in thymus from mice either untreated or 6 hours after treatment with 5 Gy whole-body IR. p53 was detected using rabbit polyclonal CM5 antibody (Leica).

Article Snippet: Cell lysates were prepared, and total cellular protein was subjected to immunoprecipitation using rabbit polyclonal anti-p53 antibody (FL393; Santa Cruz Biotechnology) followed by immunoblotting as described above using sheep polyclonal anti-p53 (AP05006PU-N; Acris Antibodies, Inc.), mouse monoclonal anti-Mdm2 (2A10, cat. #ab16895; Abcam), and mouse monoclonal anti–b-actin (cat. #A5441; Sigma).

Techniques: Mutagenesis, Isolation, Cell Culture, Control, Western Blot

Journal: Cancer Research

Article Title: The Common Germline TP53-R337H Mutation Is Hypomorphic and Confers Incomplete Penetrance and Late Tumor Onset in a Mouse Model

doi: 10.1158/0008-5472.can-20-1750

Figure Lengend Snippet: Figure 4. Mutant p53-R334H transcriptional response is attenuated in thymocytes in vivo in response to DNA damage. Affymetrix gene expression analysis of WT and R334H thymocytes either untreated (Control) or 3 hours after treatment with 5 Gy whole-body IR. Each lane represents an individual mouse. A, Heat map depicting a global perspective of all gene profiles measured by the microarray. B, Heat map display of 48 core enrichment genes in the Hallmark p53 pathway that are activated in WT cells following radiation (GSEA enrichment score 2.66, P < 0.001, FDR < 0.001). C, Heat map display of 41 core enrichment genes in the Hallmark MYC targets V2 that are activated in WT cells following radiation (FDR < 0.001). D, Quantitative real-time analysis of WT and p53-R334H thymocytes either untreated (Control) or 3 hours after treatment with 5 Gy whole-body IR (n ¼ 3).

Article Snippet: Cell lysates were prepared, and total cellular protein was subjected to immunoprecipitation using rabbit polyclonal anti-p53 antibody (FL393; Santa Cruz Biotechnology) followed by immunoblotting as described above using sheep polyclonal anti-p53 (AP05006PU-N; Acris Antibodies, Inc.), mouse monoclonal anti-Mdm2 (2A10, cat. #ab16895; Abcam), and mouse monoclonal anti–b-actin (cat. #A5441; Sigma).

Techniques: Mutagenesis, In Vivo, Gene Expression, Control, Microarray

Journal: Cancer Research

Article Title: The Common Germline TP53-R337H Mutation Is Hypomorphic and Confers Incomplete Penetrance and Late Tumor Onset in a Mouse Model

doi: 10.1158/0008-5472.can-20-1750

Figure Lengend Snippet: Figure 5. Mutant p53-R334H is functional in MEFs in response to DNA damage and in vitro senescence. A, Mutant p53-R334H confers short-term proliferative advantage in low passage MEFs. Curves represent the average of three independent MEF cell lines for each genotype counted daily in duplicate (P ¼ 0.002) B, Mutant p53-R334H competently induced cellular senescence in serially passaged MEFs (P ¼ 0.008). C, Mutant p53-R334H induces target gene expression in response to DNA damage induced by 10 mmol/L etoposide at 4 hours after treatment. Proteinswere detected using the primary antibodies p53 (PAb7); p21 (C19). D, Mutant p53-R334H induces target gene expression in response to DNA damage induced by 30 J UV radiation at 24 hours. Proteins detected using the primary antibodies p53 (1C12); p21 (F-5). MEFs at passage 4 were used in experiments presented in C and D.

Article Snippet: Cell lysates were prepared, and total cellular protein was subjected to immunoprecipitation using rabbit polyclonal anti-p53 antibody (FL393; Santa Cruz Biotechnology) followed by immunoblotting as described above using sheep polyclonal anti-p53 (AP05006PU-N; Acris Antibodies, Inc.), mouse monoclonal anti-Mdm2 (2A10, cat. #ab16895; Abcam), and mouse monoclonal anti–b-actin (cat. #A5441; Sigma).

Techniques: Mutagenesis, Functional Assay, In Vitro, Targeted Gene Expression

Journal: Cancer Research

Article Title: The Common Germline TP53-R337H Mutation Is Hypomorphic and Confers Incomplete Penetrance and Late Tumor Onset in a Mouse Model

doi: 10.1158/0008-5472.can-20-1750

Figure Lengend Snippet: Figure 6. Mutant p53-R334H has a prolonged half-life and is compromised in forming stable tetramers in primary MEFs. A, Cycloheximide half-life experiment. Early passage MEFs (passage 5) were treated with 10 mg/mL cycloheximide and harvested at indicated time points to determine p53 levels by Western blot analysis using mouse mAb 1C12. B, Protein complex analysis of primary MEFs by size exclusion chromatography and Western blot analysis (see Materials and Methods). P53 protein was detected using mAb 1C12. C, SaOS2 cells transfected with CMV Neo vector without an insert, p53 WT, or p53-R334H cDNA. Coimmunoprecipitation using Mdm2 (2A10) Ab and blotted for p53 using Sheep anti-p53 polyclonal Ab.

Article Snippet: Cell lysates were prepared, and total cellular protein was subjected to immunoprecipitation using rabbit polyclonal anti-p53 antibody (FL393; Santa Cruz Biotechnology) followed by immunoblotting as described above using sheep polyclonal anti-p53 (AP05006PU-N; Acris Antibodies, Inc.), mouse monoclonal anti-Mdm2 (2A10, cat. #ab16895; Abcam), and mouse monoclonal anti–b-actin (cat. #A5441; Sigma).

Techniques: Mutagenesis, Western Blot, Size-exclusion Chromatography, Transfection, Plasmid Preparation

Journal: Cancer Research

Article Title: The Common Germline TP53-R337H Mutation Is Hypomorphic and Confers Incomplete Penetrance and Late Tumor Onset in a Mouse Model

doi: 10.1158/0008-5472.can-20-1750

Figure Lengend Snippet: Figure 7. Mutant p53-R334H functions as a tumor suppressor in Em-Myc lympho- magenesis. A, Kaplan–Meier survival of Em-Myc transgenic WT, p53WT/R334H, and p53R334H/R334H mice. B, Western blot analysis of B-cell lymphomas aris- ing in Em-Myc WT, p53WT/R334H, and p53R334H/R334H mice. C, Characteriza- tion of p53 and p19Arf status in B-cell lymphomas arising in Em-Myc WT, p53WT/R334H, and p53R334H/R334H mice. Proteins were detected by Western blot analysis using the following prima- ry antibodies: p53 (1C12); p19Arf (ab80; ABCAM).

Article Snippet: Cell lysates were prepared, and total cellular protein was subjected to immunoprecipitation using rabbit polyclonal anti-p53 antibody (FL393; Santa Cruz Biotechnology) followed by immunoblotting as described above using sheep polyclonal anti-p53 (AP05006PU-N; Acris Antibodies, Inc.), mouse monoclonal anti-Mdm2 (2A10, cat. #ab16895; Abcam), and mouse monoclonal anti–b-actin (cat. #A5441; Sigma).

Techniques: Mutagenesis, Transgenic Assay, Western Blot

Journal: International Journal of Molecular Medicine

Article Title: lncRNA/mRNA profiling of endometriosis rat uterine tissues during the implantation window

doi: 10.3892/ijmm.2019.4370

Figure Lengend Snippet: Primers for reverse transcription-quantitative PCR.

Article Snippet: The primary antibodies against P2ry6 (1:1,000), Adamts7 (1:1,000), Dlx3 (1:1,000) and Tp53 (1:1,000; cat. no. orb319621; Biorbyt, Ltd.) were incubated at 4°C overnight.

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: lncRNA/mRNA profiling of endometriosis rat uterine tissues during the implantation window

doi: 10.3892/ijmm.2019.4370

Figure Lengend Snippet: Top 20 downregulated mRNA.

Article Snippet: The primary antibodies against P2ry6 (1:1,000), Adamts7 (1:1,000), Dlx3 (1:1,000) and Tp53 (1:1,000; cat. no. orb319621; Biorbyt, Ltd.) were incubated at 4°C overnight.

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: lncRNA/mRNA profiling of endometriosis rat uterine tissues during the implantation window

doi: 10.3892/ijmm.2019.4370

Figure Lengend Snippet: Changes in the expression of the four differentially expressed mRNAs among the three groups. mRNA expression of (A) Adamts7, (B) P2ry6 (C) Dlx3 and (D) Tp53. * P<0.05 and ** P<0.01 vs. the normal control. Adamts7, ADAM metallopeptidase with thrombospondin type 1 motif 7; Tp53, tumor protein p53; Dlx3, distal-less homeobox 3; P2ry6, pyrimidinergic receptor P2Y6.

Article Snippet: The primary antibodies against P2ry6 (1:1,000), Adamts7 (1:1,000), Dlx3 (1:1,000) and Tp53 (1:1,000; cat. no. orb319621; Biorbyt, Ltd.) were incubated at 4°C overnight.

Techniques: Expressing, Control

Journal: International Journal of Molecular Medicine

Article Title: lncRNA/mRNA profiling of endometriosis rat uterine tissues during the implantation window

doi: 10.3892/ijmm.2019.4370

Figure Lengend Snippet: Immunohistochemistry of uterine tissue in rats. (A) Expression of Adamts7, P2ry6, Dlx3 and Tp53 in the endometriosis group (n=13), adipose tissue control group (n=8) and blank control group (n=14). (magnification, x200). Quantification of the immunohistochemistry results for (B) Adamts7, (C) P2ry6, (D) Dlx3 and (E) Tp53. The samples were tested in triplicates. * P<0.05 vs. blank controls. # P<0.05 vs. adipose tissue controls. Adamts7, ADAM metallopep-tidase with thrombospondin type 1 motif 7; Tp53, tumor protein p53; Dlx3, distal-less homeobox 3; P2ry6, pyrimidinergic receptor P2Y6; GE, glandular epithelium; St, endometrial stroma; UE, uterine epithelium.

Article Snippet: The primary antibodies against P2ry6 (1:1,000), Adamts7 (1:1,000), Dlx3 (1:1,000) and Tp53 (1:1,000; cat. no. orb319621; Biorbyt, Ltd.) were incubated at 4°C overnight.

Techniques: Immunohistochemistry, Expressing, Control

Journal: International Journal of Molecular Medicine

Article Title: lncRNA/mRNA profiling of endometriosis rat uterine tissues during the implantation window

doi: 10.3892/ijmm.2019.4370

Figure Lengend Snippet: Western blot analysis of uterine tissue in rats. (A) Expression of Adamts7, P2ry6, Dlx3 and Tp53 in the endometriosis group (n=13), adipose tissue control group (n=8) and blank control group (n=14). Quantification of the western blotting results for (B) Adamts7, (C) P2ry6, (D) Dlx3 and (E) Tp53. The samples were tested in triplicates. β-actin was used to normalize the data. ** P<0.01 vs. normal control, ## P<0.01 vs. adipose tissue controls. Adamts7, ADAM metallopeptidase with thrombospondin type 1 motif 7; Tp53, tumor protein p53; Dlx3, distal-less homeobox 3; P2ry6, pyrimidinergic receptor P2Y6.

Article Snippet: The primary antibodies against P2ry6 (1:1,000), Adamts7 (1:1,000), Dlx3 (1:1,000) and Tp53 (1:1,000; cat. no. orb319621; Biorbyt, Ltd.) were incubated at 4°C overnight.

Techniques: Western Blot, Expressing, Control

Journal: International Journal of Molecular Medicine

Article Title: lncRNA/mRNA profiling of endometriosis rat uterine tissues during the implantation window

doi: 10.3892/ijmm.2019.4370

Figure Lengend Snippet: Expression of lncRNA and mRNA.

Article Snippet: The primary antibodies against P2ry6 (1:1,000), Adamts7 (1:1,000), Dlx3 (1:1,000) and Tp53 (1:1,000; cat. no. orb319621; Biorbyt, Ltd.) were incubated at 4°C overnight.

Techniques: Expressing

Journal: International Journal of Molecular Medicine

Article Title: lncRNA/mRNA profiling of endometriosis rat uterine tissues during the implantation window

doi: 10.3892/ijmm.2019.4370

Figure Lengend Snippet: Relationships among the five differentially expressed lncRNAs and the four mRNAs. The lncRNAs and mRNA shown here are those that were validated by quantitative PCR. The samples were tested in triplicates. lnc, long noncoding; Adamts7, ADAM metallopeptidase with thrombospondin type 1 motif 7; Tp53, tumor protein p53; Dlx3, distal-less homeobox 3; P2ry6, pyrimidinergic receptor P2Y6.

Article Snippet: The primary antibodies against P2ry6 (1:1,000), Adamts7 (1:1,000), Dlx3 (1:1,000) and Tp53 (1:1,000; cat. no. orb319621; Biorbyt, Ltd.) were incubated at 4°C overnight.

Techniques: Real-time Polymerase Chain Reaction

Journal: Cancer Medicine

Article Title: The GNL3L ‐ MDM2 Interaction Drives Esophageal Squamous Cell Carcinoma Progression

doi: 10.1002/cam4.71146

Figure Lengend Snippet: GNL3L positively regulated MDM2 expression and interacted with MDM2. (a) After GNL3L knockdown, qRT‐PCR, and western blot (WB) were performed to determine MDM2 expression in TE‐1 cells. (b) Differences in the expression of p53 and p21 in TE‐1 cells after the knockdown of GNL3L. (c) Co‐IP of GNL3L and MDM2 from ESCC cell lines. The input represents the total protein extract used in the IP. The GNL3L protein expression was normalized to that of GAPDH in ESCC cells. (IP, immunoprecipitation; IgG, negative control). (d) qRT‐PCR analysis of MDM2 and GNL3L expression in TE‐1 cells in different groups. (e) Protein expression of MDM2 and GNL3L of TE‐1 in different groups was determined by WB. (f) Cell proliferation was analyzed in different groups using the CCK8 assay for TE‐1 cell lines. (g) Proliferation was analyzed using an EdU assay. (** p < 0.01, ## p < 0.01).

Article Snippet: The primary antibodies were GAPDH polyclonal antibody (1:3000, Affinity, AF7021, 36 kDa), MDM2 polyclonal antibody (1:1000, Affinity, AF0208, 90 kDa), GNL3L antibody (1:1000, Affinity, DF4113, 66 kDa), p53 antibody (1:1000, Affinity, AF0879, 53 kDa), and p21 antibody (1:1000, MedChemExpress, HY‐ P80774 , 18 kDa).

Techniques: Expressing, Knockdown, Quantitative RT-PCR, Western Blot, Co-Immunoprecipitation Assay, Immunoprecipitation, Negative Control, CCK-8 Assay, EdU Assay

Journal: Cancer Medicine

Article Title: The GNL3L ‐ MDM2 Interaction Drives Esophageal Squamous Cell Carcinoma Progression

doi: 10.1002/cam4.71146

Figure Lengend Snippet: GNL3L knockdown reversed the increased malignant behaviors of ESCC cells induced by MDM2 overexpression. (a) Wound healing assay results in TE‐1 cells from different groups. (b) Invasion and migration of TE‐1 cells were assessed using Transwell assays. (c) Distribution of the cell phases of TE‐1 in the different groups. (d) In the different groups, apoptosis was measured in TE‐1 cells using flow cytometry. (e) Expression of P53 and P21 in TE‐1 cells in different groups. (** p < 0.01, ## p < 0.01).

Article Snippet: The primary antibodies were GAPDH polyclonal antibody (1:3000, Affinity, AF7021, 36 kDa), MDM2 polyclonal antibody (1:1000, Affinity, AF0208, 90 kDa), GNL3L antibody (1:1000, Affinity, DF4113, 66 kDa), p53 antibody (1:1000, Affinity, AF0879, 53 kDa), and p21 antibody (1:1000, MedChemExpress, HY‐ P80774 , 18 kDa).

Techniques: Knockdown, Over Expression, Wound Healing Assay, Migration, Flow Cytometry, Expressing

Journal: Cancer Medicine

Article Title: The GNL3L ‐ MDM2 Interaction Drives Esophageal Squamous Cell Carcinoma Progression

doi: 10.1002/cam4.71146

Figure Lengend Snippet: GNL3L overexpression reversed the decreased malignant behavior of ESCC cells induced by MDM2 knockdown. (a) Protein expression of MDM2 and GNL3L in TE‐1 cells in different groups was determined by western blot (WB). (b) Cell proliferation was analyzed in different groups using an EdU assay. (c) Migration and invasion of TE‐1 cells were assessed using Transwell assays. (d) In the different groups, apoptosis was measured in TE‐1 cells using flow cytometry. (e) Expression of P53 and P21 in TE‐1 cells in different groups. (** p < 0.01, ## p < 0.01).

Article Snippet: The primary antibodies were GAPDH polyclonal antibody (1:3000, Affinity, AF7021, 36 kDa), MDM2 polyclonal antibody (1:1000, Affinity, AF0208, 90 kDa), GNL3L antibody (1:1000, Affinity, DF4113, 66 kDa), p53 antibody (1:1000, Affinity, AF0879, 53 kDa), and p21 antibody (1:1000, MedChemExpress, HY‐ P80774 , 18 kDa).

Techniques: Over Expression, Knockdown, Expressing, Western Blot, EdU Assay, Migration, Flow Cytometry