rabbit polyclonal anti fdps  (Proteintech)

Journal: Cells

Article Title: Cholesterol Homeostasis Modulates Platinum Sensitivity in Human Ovarian Cancer

doi: 10.3390/cells9040828

Figure Lengend Snippet: Cholesterol synthesis inhibition increases chemoresistance in platinum-sensitive OC cells. ( A ) PEA1, PEA2, and PEO14 cells were treated with 5 μM Lovastatin and then treated with 20 μM cisplatin for an additional 48 h (when medium was changed, Lovastatin was added again). Subsequently, apoptosis was measured by a luminescent caspase 3/7 activity assay. Data are expressed as mean ± S.E.M. from four (PEA1 and PEA2) or six (PEO14) independent experiments with technical triplicates each. The numbers above the bars indicate the statistical significance ( p -value), based on the two-tailed unpaired Student’s t -test (significant values highlighted in red). ( B ) Twenty-four hours after seeding PEA1 and PEA2 cells in standard culture medium, cells were cultured for additional 48 or 96 h in medium supplemented with lipid-stripped serum. Finally, cell viability was measured by an MTT-based cell viability assay. ( C ) Immunoblot of total lysates from PEA1 and PEA2 cells 48 h after lipid withdrawal from culture medium. Images are representative of five independent experiments. Bar plots below panels show mean ± S.E.M. of relative densitometric band intensity calculated by assuming each band of the control condition (normalized on Actin) = 1 ( n = 5). The numbers above the bars represent the statistical significance ( p -value) based on the Student’s t -test (significant values highlighted in red). ( D ) PEA2 platinum-resistant cells were cultured for 48 h in medium supplemented with lipid-stripped serum and then treated with 40 μM cisplatin for additional 48 h. Apoptosis was measured by a luminescent caspase 3/7 activity assay. Data are expressed as mean ± S.E.M. from four independent experiments with technical triplicates each. The numbers above the bars indicate the statistical significance ( p -value), based on the two-tailed unpaired Student’s t -test. ( E , F ) Kaplan–Meier estimates of the impact of FDPS ( E ) and LDLR ( F ) on overall survival in OC, according to The Cancer Genome Atlas (TCGA) database.

Article Snippet: The following antibodies were used: anti-TRAP1 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA; sc-13557), anti-β-Actin (Santa Cruz Biotechnology, Inc., Dallas, TX, USA; sc-69879), anti-LDLR (Santa Cruz Biotechnology, Inc., Dallas, TX, USA; sc-18823), anti-FDPS (OriGene, Herford, Germany; AP12197PU-N), and anti-OSC (Santa Cruz Biotechnology, Inc., Dallas, TX, USA; sc-514507).

Techniques: Inhibition, Activity Assay, Two Tailed Test, Cell Culture, Viability Assay, Western Blot

Journal: Cells

Article Title: Cholesterol Homeostasis Modulates Platinum Sensitivity in Human Ovarian Cancer

doi: 10.3390/cells9040828

Figure Lengend Snippet: A proposed model for the remodeling of cholesterol metabolism in the transition from a chemosensitive to a chemoresistant phenotype in human ovarian cancer cells. Drug-sensitive cells show significant activation of the cholesterol biosynthetic pathway and transport of cholesterol through the Golgi apparatus, while uptake of exogenous cholesterol through the LDLR is limited. Drug resistant cells, on the opposite, show reduced levels of FDPS and OSC along the biosynthetic pathway and increased LDL uptake. The inhibition of 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR), the rate-limiting enzyme of the biosynthetic pathway, by statins induces drug resistance, whereas the reduction of LDL recycling and lipid absorption through extracellular lipid withdrawal or LIPG knockdown sensitizes cells to drug-induced apoptosis. Enzymes whose expression has been analyzed in this work are in bold red. IPP = isopentenyl pyrophosphate; FPP = farnesyl pyrophosphate; TCA = tricarboxylic acid. This image was created using images from Servier Medical Art under Creative Commons Attribution 3.0 Unported License ( https://smart.servier.com ).

Article Snippet: The following antibodies were used: anti-TRAP1 (Santa Cruz Biotechnology, Inc., Dallas, TX, USA; sc-13557), anti-β-Actin (Santa Cruz Biotechnology, Inc., Dallas, TX, USA; sc-69879), anti-LDLR (Santa Cruz Biotechnology, Inc., Dallas, TX, USA; sc-18823), anti-FDPS (OriGene, Herford, Germany; AP12197PU-N), and anti-OSC (Santa Cruz Biotechnology, Inc., Dallas, TX, USA; sc-514507).

Techniques: Activation Assay, Inhibition, Expressing

Journal: BioMed Research International

Article Title: Lentiviral-Mediated Silencing of Farnesyl Pyrophosphate Synthase through RNA Interference in Mice

doi: 10.1155/2015/914026

Figure Lengend Snippet: Sequences of five siRNAs targeting murine FPPS and one nontargeting control.

Article Snippet: The membranes were blocked with 5% skim milk in Tween/TBS (TBST) for 1 h and subsequently incubated overnight at 4°C with rabbit polyclonal anti-FPPS antibody (ProteinTech) (1 : 1500 dilution) or mouse monoclonal anti- β -actin antibody (GenScript) (1 : 2,000 dilution).

Techniques: Control, Sequencing

Journal: BioMed Research International

Article Title: Lentiviral-Mediated Silencing of Farnesyl Pyrophosphate Synthase through RNA Interference in Mice

doi: 10.1155/2015/914026

Figure Lengend Snippet: Sequences of primers used for real-time polymerase chain reaction.

Article Snippet: The membranes were blocked with 5% skim milk in Tween/TBS (TBST) for 1 h and subsequently incubated overnight at 4°C with rabbit polyclonal anti-FPPS antibody (ProteinTech) (1 : 1500 dilution) or mouse monoclonal anti- β -actin antibody (GenScript) (1 : 2,000 dilution).

Techniques: Sequencing

Journal: BioMed Research International

Article Title: Lentiviral-Mediated Silencing of Farnesyl Pyrophosphate Synthase through RNA Interference in Mice

doi: 10.1155/2015/914026

Figure Lengend Snippet: (a) Western blot data of FPPS expression in cardiomyocytes infected with different lentivirus. (b) Relative expression levels of FPPS normalized to β -actin are expressed as the mean ± SEM. Identical results were obtained in three independent experiments. ** P < 0.01 versus pLVT7 (nontargeting control).

Article Snippet: The membranes were blocked with 5% skim milk in Tween/TBS (TBST) for 1 h and subsequently incubated overnight at 4°C with rabbit polyclonal anti-FPPS antibody (ProteinTech) (1 : 1500 dilution) or mouse monoclonal anti- β -actin antibody (GenScript) (1 : 2,000 dilution).

Techniques: Western Blot, Expressing, Infection, Control

Journal: BioMed Research International

Article Title: Lentiviral-Mediated Silencing of Farnesyl Pyrophosphate Synthase through RNA Interference in Mice

doi: 10.1155/2015/914026

Figure Lengend Snippet: (a) Western blot data of FPPS expression in transgenic mice. Left: samples of each tissue were from nontransgenic mice and those on the right were from transgenic mice. (b) Relative expression levels of FPPS normalized to β -actin are expressed as the mean ± SEM ( n = 3). NTg: non-transgenic; Tg: transgenic (F2). * P < 0.05; ** P < 0.01 versus NTg mice.

Article Snippet: The membranes were blocked with 5% skim milk in Tween/TBS (TBST) for 1 h and subsequently incubated overnight at 4°C with rabbit polyclonal anti-FPPS antibody (ProteinTech) (1 : 1500 dilution) or mouse monoclonal anti- β -actin antibody (GenScript) (1 : 2,000 dilution).

Techniques: Western Blot, Expressing, Transgenic Assay

Journal: BioMed Research International

Article Title: Lentiviral-Mediated Silencing of Farnesyl Pyrophosphate Synthase through RNA Interference in Mice

doi: 10.1155/2015/914026

Figure Lengend Snippet: FPPS expression in transgenic mice. (a) Relative mRNA expression levels of FPPS normalized to β -actin are expressed as the mean ± SEM ( n = 3). NTg: nontransgenic; Tg: transgenic (F2). * P < 0.05; ** P < 0.01; *** P < 0.001 versus NTg mice. (b) FPPS protein expression in F2 and F3 mice. Left: samples of heart tissue were from nontransgenic mice and those on the right were from transgenic F2 and F3 mice.

Article Snippet: The membranes were blocked with 5% skim milk in Tween/TBS (TBST) for 1 h and subsequently incubated overnight at 4°C with rabbit polyclonal anti-FPPS antibody (ProteinTech) (1 : 1500 dilution) or mouse monoclonal anti- β -actin antibody (GenScript) (1 : 2,000 dilution).

Techniques: Expressing, Transgenic Assay