Journal: The Journal of biological chemistry

Article Title: Modulation of interleukin-6-induced plasma protein secretion in hepatoma cells by p53 species.

doi: 10.1074/jbc.270.39.23159

Figure Lengend Snippet: FIG. 2. Modulation by p53 species of IL-6-induced activation of pbFibCAT in hepatoma cells. Cultures of Hep3B (panel A) or HepG2 (panels B and C) cells in 100-mm plastic dishes cotransfected with the enhancer/reporter pbFibCAT (10 mg) and various p53 expression vec- tors (5 mg) were treated with IL-6 (30 ng/ml) for 24 h and the level of CAT expression monitored (bgal normalized). The level of CAT expres- sion is depicted as fold change (6 standard deviation) with reference to the CAT activity in cells not treated with IL-6 or any p53 vector taken as 1. n represents the number of replications of each test combination; statistical evaluation was carried out using the Student’s two-tailed t test. Panel C is an autoradiogram that illustrates CAT assays corre- sponding to one experiment. p53 constructs: SN3, human wt; CX3, human mutant; Nc9, murine wt; c5, Val135, and Phe132, murine mutants.

Article Snippet: The cultures were then washed with cold phosphate-buffered saline, fixed with chilled acetone for 2 min, and air-dried. p53 antigen was detected using the mouse-monoclonal antibody Pab240 (pan-reactive to human and murine wt and mutant p53 species) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) as described by the manufacturer.

Techniques: Activation Assay, Expressing, Standard Deviation, Activity Assay, Plasmid Preparation, Two Tailed Test, Construct, Mutagenesis

Journal: The Journal of biological chemistry

Article Title: Modulation of interleukin-6-induced plasma protein secretion in hepatoma cells by p53 species.

doi: 10.1074/jbc.270.39.23159

Figure Lengend Snippet: FIG. 3. Modulation by p53 species of the muscle-specific crea- tine kinase-derived enhancer/reporter p50–2 in IL-6-treated hepatoma cells. Duplicate cultures of Hep3B cells (panels A and B) in 100-mm plastic dishes cotransfected with the enhancer/reporter p50–2 (10 mg) and various p53 expression vectors (5 mg) were treated with IL-6 (30 ng/ml) for 24 h and the CAT expression monitored essentially as described in legend to Fig. 2. p53 constructs: SN3, human wt; CX3, human mutant; Nc9, murine wt; c5, Val135, and Phe132, murine mu- tants.

Article Snippet: The cultures were then washed with cold phosphate-buffered saline, fixed with chilled acetone for 2 min, and air-dried. p53 antigen was detected using the mouse-monoclonal antibody Pab240 (pan-reactive to human and murine wt and mutant p53 species) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) as described by the manufacturer.

Techniques: Derivative Assay, Expressing, Construct, Mutagenesis

Journal: The Journal of biological chemistry

Article Title: Modulation of interleukin-6-induced plasma protein secretion in hepatoma cells by p53 species.

doi: 10.1074/jbc.270.39.23159

Figure Lengend Snippet: FIG. 4. Modulation by p53 species of C/EBP-induced activation of pbFibCAT in hepatoma cells. HepG2 cultures in 100-mm dishes were cotransfected with pbFibCAT (10 mg), expression vectors for C/EBPa, b, or d (5 mg) together with various p53 expression vectors (5 mg) in duplicate. Cells were harvested 24 h after the beginning of transfection and the level of CAT expression monitored essentially as described in legend to Fig. 2. Inset in panel C shows an autoradiogram of a CAT assay from one experiment. p53 constructs: SN3, human wt; CX3, human mutant; Nc9, murine wt; c5, Val135, and Phe132, murine mutants.

Article Snippet: The cultures were then washed with cold phosphate-buffered saline, fixed with chilled acetone for 2 min, and air-dried. p53 antigen was detected using the mouse-monoclonal antibody Pab240 (pan-reactive to human and murine wt and mutant p53 species) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) as described by the manufacturer.

Techniques: Activation Assay, Expressing, Transfection, Construct, Mutagenesis

Journal: The Journal of biological chemistry

Article Title: Modulation of interleukin-6-induced plasma protein secretion in hepatoma cells by p53 species.

doi: 10.1074/jbc.270.39.23159

Figure Lengend Snippet: FIG. 5. p53 immunostaining in hepatoma cell lines. Parental Hep3B cells (panel A), three p53Val-1351-expressing hepatoma lines (lines 1–3, respectively, panels B-D), and a hepatoma line transfected with pSVneo alone (line 22, panel D) were tested for p53 by immuno- staining using a panreactive anti-p53 monoclonal antibody (Pab 240) (reactive with wt and mutant human and murine p53 species). The length marker equals 20 mm.

Article Snippet: The cultures were then washed with cold phosphate-buffered saline, fixed with chilled acetone for 2 min, and air-dried. p53 antigen was detected using the mouse-monoclonal antibody Pab240 (pan-reactive to human and murine wt and mutant p53 species) (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) as described by the manufacturer.

Techniques: Immunostaining, Expressing, Transfection, Mutagenesis, Marker

Journal: PloS one

Article Title: The anti-hypercholesterolemic effect of low p53 expression protects vascular endothelial function in mice.

doi: 10.1371/journal.pone.0092394

Figure Lengend Snippet: Figure 1. p53 protein expression and its target gene p21 mRNA expression. A) liver and B) aorta of WT and p53+/2 6-month old mice. a-actin for aorta and b-actin for liver were used as internal loading controls. Data are mean6SEM ; n = 5–9. #p,0.05. doi:10.1371/journal.pone.0092394.g001

Article Snippet: Animals Wild type C57Bl/6J (WT; B6/129PF2/J of the following parental strains: C57BL/6J-Aw–J and 129P3/J (formerly 129/J)) and B6.129S2-Trp53tm1Tyj/J heterozygous (p53+/2) [17] 3-monthold male mice were purchased from The Jackson Laboratory (Bar Harbor, Maine, USA).

Techniques: Expressing

Journal: PloS one

Article Title: The anti-hypercholesterolemic effect of low p53 expression protects vascular endothelial function in mice.

doi: 10.1371/journal.pone.0092394

Figure Lengend Snippet: Figure 2. Similar synthesis and uptake of cholesterol. Hepatic mRNA expression of important genes implicated in cholesterol metabolism in WT and p53+/2 6-month old mice fed a regular diet (RD) or a western diet (WD). A) HMG CoA reductase, B) LDL receptor (LDLR), C) apolipoprotein B (apoB) and D) proprotein convertase subtilisin kexin type 9 (PCSK9). Data are mean6SEM ; n = 5–7. doi:10.1371/journal.pone.0092394.g002

Article Snippet: Animals Wild type C57Bl/6J (WT; B6/129PF2/J of the following parental strains: C57BL/6J-Aw–J and 129P3/J (formerly 129/J)) and B6.129S2-Trp53tm1Tyj/J heterozygous (p53+/2) [17] 3-monthold male mice were purchased from The Jackson Laboratory (Bar Harbor, Maine, USA).

Techniques: Expressing, Western Blot

Journal: PloS one

Article Title: The anti-hypercholesterolemic effect of low p53 expression protects vascular endothelial function in mice.

doi: 10.1371/journal.pone.0092394

Figure Lengend Snippet: Figure 3. Key mediators of bile acids (BA) synthesis in the liver of mice. A) Transcription factors p53 and farnesoid X receptor (FXR) can activate the transcription of small heterodimer partner (SHP) which inhibits 7-a-hydroxylase (Cyp7A1), the first enzyme in the BA synthesis pathway. Liver X receptor (LXR) can activate transcriptionally Cyp7A1 to produce more BA. B) The Western diet (WD) activated transcription of LXR in WT mice in association with increased Cyp7A1 gene expression. Lower p53 expression in p53+/2 mice is associated with lower gene expression of SHP. While no difference is seen in FXR and LXR, Cyp7A1 gene expression and BA levels are elevated. doi:10.1371/journal.pone.0092394.g003

Article Snippet: Animals Wild type C57Bl/6J (WT; B6/129PF2/J of the following parental strains: C57BL/6J-Aw–J and 129P3/J (formerly 129/J)) and B6.129S2-Trp53tm1Tyj/J heterozygous (p53+/2) [17] 3-monthold male mice were purchased from The Jackson Laboratory (Bar Harbor, Maine, USA).

Techniques: Western Blot, Gene Expression, Expressing

Journal: PloS one

Article Title: The anti-hypercholesterolemic effect of low p53 expression protects vascular endothelial function in mice.

doi: 10.1371/journal.pone.0092394

Figure Lengend Snippet: Figure 4. Bile acid genes expression differs between p53+/2 and WT mice. Hepatic mRNA expression of genes implicated in bile acid metabolism in WT and p53+/2 6-month-old mice fed a regular diet (RD) or a 3-month Western diet (WD). A) 7-alpha-hydroxylase (Cyp7A1), B) liver X receptors (LXR), C) small heterodimer partner (SHP), and D) farnesoid X receptors (FXR). Data are means6SEM; n = 5–7. *p,0.05 compared to RD with matching genotype; #p,0.05 compared to WT mice with matching diet. doi:10.1371/journal.pone.0092394.g004

Article Snippet: Animals Wild type C57Bl/6J (WT; B6/129PF2/J of the following parental strains: C57BL/6J-Aw–J and 129P3/J (formerly 129/J)) and B6.129S2-Trp53tm1Tyj/J heterozygous (p53+/2) [17] 3-monthold male mice were purchased from The Jackson Laboratory (Bar Harbor, Maine, USA).

Techniques: Expressing, Western Blot

Journal: PloS one

Article Title: The anti-hypercholesterolemic effect of low p53 expression protects vascular endothelial function in mice.

doi: 10.1371/journal.pone.0092394

Figure Lengend Snippet: Figure 5. Effect of doxorubicin (DOX) on gene expression in HepG2 cells. Gene expression of A) p53, B) p21, C) SHP and D) Cyp7A1 in human hepatic HepG2 cell lines after a 24-hour incubation with 0.5 mg/ml DOX, a known-activator of p53. Data are mean6SEM of four independent experiments analyzed in duplicate. doi:10.1371/journal.pone.0092394.g005

Article Snippet: Animals Wild type C57Bl/6J (WT; B6/129PF2/J of the following parental strains: C57BL/6J-Aw–J and 129P3/J (formerly 129/J)) and B6.129S2-Trp53tm1Tyj/J heterozygous (p53+/2) [17] 3-monthold male mice were purchased from The Jackson Laboratory (Bar Harbor, Maine, USA).

Techniques: Gene Expression, Incubation

Journal: PloS one

Article Title: The anti-hypercholesterolemic effect of low p53 expression protects vascular endothelial function in mice.

doi: 10.1371/journal.pone.0092394

Figure Lengend Snippet: Figure 6. Age and WD affect differentially p53+/2 and WT mice. Aortic endothelium-dependent relaxation concentration-response curves with acetylcholine (Ach) in 3-month (3-mo) and 6-month (6-mo) WT or p53+/2 mice fed a regular diet (RD) or Western diet (WD). Data are means6SEM; n = 4–14. &p,0.05 compared to 3-mo; *p,0.05 compared to RD; #p,0.05 compared to 6-mo WT RD mice. doi:10.1371/journal.pone.0092394.g006

Article Snippet: Animals Wild type C57Bl/6J (WT; B6/129PF2/J of the following parental strains: C57BL/6J-Aw–J and 129P3/J (formerly 129/J)) and B6.129S2-Trp53tm1Tyj/J heterozygous (p53+/2) [17] 3-monthold male mice were purchased from The Jackson Laboratory (Bar Harbor, Maine, USA).

Techniques: Concentration Assay, Western Blot

Journal: PloS one

Article Title: The anti-hypercholesterolemic effect of low p53 expression protects vascular endothelial function in mice.

doi: 10.1371/journal.pone.0092394

Figure Lengend Snippet: Figure 7. Effect at 6-month on thromboxane A2 pathway. Measures in WT and p53+/2 mice fed a regular diet (RD) of A) endothelium- dependent relaxations to acetylcholine (ACh) of aorta segments isolated from WT and p53+/2 mice were obtained in presence of thromboxane synthase inhibition (furegrelate, 10 mM), B) thromboxane synthase (TXS) gene expression in the aorta, C) release of the thromboxane A2 metabolite 11-dehydro thromboxane B2 (TXB2) in the plasma. Data are mean6SEM. n = 3–6 in (A), n = 3 in (B) and n = 4 in (C). doi:10.1371/journal.pone.0092394.g007

Article Snippet: Animals Wild type C57Bl/6J (WT; B6/129PF2/J of the following parental strains: C57BL/6J-Aw–J and 129P3/J (formerly 129/J)) and B6.129S2-Trp53tm1Tyj/J heterozygous (p53+/2) [17] 3-monthold male mice were purchased from The Jackson Laboratory (Bar Harbor, Maine, USA).

Techniques: Isolation, Inhibition, Gene Expression, Clinical Proteomics

Journal: PloS one

Article Title: The anti-hypercholesterolemic effect of low p53 expression protects vascular endothelial function in mice.

doi: 10.1371/journal.pone.0092394

Figure Lengend Snippet: Figure 8. Adaptive endothelium-dependent mechanisms are not activated in p53+/2 mice. Effect of WD on non-NO/non-prostacyclin endothelium-dependent relaxation of aorta from WT and p53+/2 mice. Endothelium-dependent relaxation by acetylcholine (Ach) of aortic segments from WT and p53+/2 mice was assessed in the presence of NOS inhibition (L-NNA, 100 mM) and nonspecific COX1/2 inhibition (indomethacin, 10 mM). Data are means6SEM. n = 5–10. doi:10.1371/journal.pone.0092394.g008

Article Snippet: Animals Wild type C57Bl/6J (WT; B6/129PF2/J of the following parental strains: C57BL/6J-Aw–J and 129P3/J (formerly 129/J)) and B6.129S2-Trp53tm1Tyj/J heterozygous (p53+/2) [17] 3-monthold male mice were purchased from The Jackson Laboratory (Bar Harbor, Maine, USA).

Techniques: Inhibition

Journal: PloS one

Article Title: The anti-hypercholesterolemic effect of low p53 expression protects vascular endothelial function in mice.

doi: 10.1371/journal.pone.0092394

Figure Lengend Snippet: Figure 9. Aortic oxidative stress level and SOD2 activity. (A) Staining with dihydroethidium (DHE, red) of WT and p53+/2 6-month old mice fed a regular diet (RD) or a 3-month western diet (WD). Data are mean6SEM; n = 3–4. (B) Effect of the WD on superoxide dismutase 2 (SOD2) activity in aortic segments from WT and p53+/2 mice. Data are mean6SEM; n = 3–8. *p,0.05 compared to RD with matching genotype. doi:10.1371/journal.pone.0092394.g009

Article Snippet: Animals Wild type C57Bl/6J (WT; B6/129PF2/J of the following parental strains: C57BL/6J-Aw–J and 129P3/J (formerly 129/J)) and B6.129S2-Trp53tm1Tyj/J heterozygous (p53+/2) [17] 3-monthold male mice were purchased from The Jackson Laboratory (Bar Harbor, Maine, USA).

Techniques: Activity Assay, Staining, Western Blot

Journal: Cancer research

Article Title: Concurrent inhibition of IGF1R and ERK increases pancreatic cancer sensitivity to autophagy inhibitors

doi: 10.1158/0008-5472.CAN-21-1443

Figure Lengend Snippet: Dual inhibition of IGF1R and autophagy impairs PDAC proliferation in 3D and in vivo models. A, Human (Pa01C, Pa14C) and murine (mT4) PDAC cell lines were grown as spheroids and treated with or without BMS-754807 (IGF1Ri, 200 nM) and/or CQ (6 μM) for 72 hours and cytotoxicity was assayed via the addition of the apoptotic dye CellTox Green (500 nM). Representative images at 0 and 72 hours of drug treatment are shown (top). Scale bar = 400 μm. Total integrated intensity of CellTox fluorescence at 72 hours after treatment is plotted (bottom). Integrated CellTox fluorescence intensity is representative of at least four independent experiments. B, Ex vivo tumor weight of mT4 mouse xenografts following treatment with vehicle, BMS-754807 (IGF1Ri, 12.5 mg/kg), HCQ (60 mg/kg), and the combination. C, GSEA of Hallmark gene sets following Clariom S Assay HT mouse microarray of mRNA prepared from six representative mT4 tumors per condition from B .

Article Snippet: The mouse tumor-derived Kras -mutant cell line mT4 [KPC ( Kras G12D/+ ; Tp53 R172H/+ ; Pdx-Cre)] and the KRAS WT hF39 and hF43 cell lines were provided by D. Tuveson (Cold Spring Harbor Laboratory, NY).

Techniques: Inhibition, In Vivo, CellTox Assay, Fluorescence, Ex Vivo, Microarray