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    Horizon Discovery raf1 sirna
    cells were placed in suspension, or adherent cultures were serum-starved for two hours seventy-two hours post transfection with control or <t>Raf1</t> <t>siRNA.</t> Adherent cultures were then stimulated with EGF (1 ng/mL for 5 min) or plated on fibronectin (FN)-coated dishes for the indicated times. Cell lysates were analyzed by western blot with the indicated antisera.
    Raf1 Sirna, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery rat raf 1 selective sirna mixture
    Intrathecal <t>Raf-1-selective</t> <t>siRNA</t> pretreatment attenuates sustained morphine-mediated tactile allodynia. For description of the animal groups see . Paw withdrawal thresholds in response to a series of von Frey filaments applied to the plantar surface
    Rat Raf 1 Selective Sirna Mixture, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Horizon Discovery lipid encapsulated raf 1 selective sirna mixture
    A. Experimental design. (Ith: Intrathecal; <t>siRNA:</t> <t>Raf-1</t> siRNA or mismatch siRNA delivery via intrathecal catheter; BL: Baseline; D: Day; IR: Infrared heat). B. Raf-1 siRNA attenuates sustained morphine-mediated thermal hyperalgesia in rats: Male Sprague Dawley rats received intrathecal injections of i-Fect encapsulated Raf-1selective siRNA (Raf-1 siRNA groups); or non-targeting dsRNA (mismatch siRNA groups) or the transfection agent (i-Fect) alone (control) once daily, for 3 days. After pre-treatment, the rats received continous subcutaneous (osmotic minipump) saline (control group, Raf-1 siRNA group and mismatch siRNA group) or morphine (45μg/μl/h) (morphine group, Raf-1 siRNA+morphine group and mismatch siRNA+morphine group) infusions for 6 days. Six animals were included in each treatment group. Thermal hyperalgesia was measured as a decrease in paw withdrawal latencies in a radiant heat paw-withdrawal test.
    Lipid Encapsulated Raf 1 Selective Sirna Mixture, supplied by Horizon Discovery, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher raf 1 small inhibitory rna sirna
    <t>Raf-1</t> is essential for ERK activation by EGF and for PAK165-dependent rescue of ERK activation in cells treated with forskolin and IBMX. (A) Effects of Raf-1 <t>siRNA.</t> HEK293 cells were transfected with GFPERK, vector (−), or PAK165 (+) and
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    Santa Cruz Biotechnology raf 1
    RRD-251 inhibits <t>Rb-Raf-1</t> interaction in AoSMCs. (A and B) Treatment with TNFα or PDGF in the presence of RRD-251 inhibits Raf-1 levels in AoSMCs (A) and A10s (B). (C) TNFα and PDGF treatment induced Rb-Raf-1 binding in AoSMCs. (D) RRD-251 inhibits TNFα-induced Rb-Raf-1 interaction.
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    Horizon Discovery smartpool sirna against raf1 siraf
    RRD-251 inhibits <t>Rb-Raf-1</t> interaction in AoSMCs. (A and B) Treatment with TNFα or PDGF in the presence of RRD-251 inhibits Raf-1 levels in AoSMCs (A) and A10s (B). (C) TNFα and PDGF treatment induced Rb-Raf-1 binding in AoSMCs. (D) RRD-251 inhibits TNFα-induced Rb-Raf-1 interaction.
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    Image Search Results


    cells were placed in suspension, or adherent cultures were serum-starved for two hours seventy-two hours post transfection with control or Raf1 siRNA. Adherent cultures were then stimulated with EGF (1 ng/mL for 5 min) or plated on fibronectin (FN)-coated dishes for the indicated times. Cell lysates were analyzed by western blot with the indicated antisera.

    Journal: Cellular signalling

    Article Title: MEK1 activation by PAK: A novel mechanism

    doi: 10.1016/j.cellsig.2007.01.018

    Figure Lengend Snippet: cells were placed in suspension, or adherent cultures were serum-starved for two hours seventy-two hours post transfection with control or Raf1 siRNA. Adherent cultures were then stimulated with EGF (1 ng/mL for 5 min) or plated on fibronectin (FN)-coated dishes for the indicated times. Cell lysates were analyzed by western blot with the indicated antisera.

    Article Snippet: Cells were transfected with 100 nM control or Raf1 siRNA (SMARTpool, Dharmacon) using 30 μL Lipofectamine 2000 (Invitrogen).

    Techniques: Transfection, Western Blot

    Intrathecal Raf-1-selective siRNA pretreatment attenuates sustained morphine-mediated tactile allodynia. For description of the animal groups see . Paw withdrawal thresholds in response to a series of von Frey filaments applied to the plantar surface

    Journal: British Journal of Pharmacology

    Article Title: Sustained morphine-mediated pain sensitization and antinociceptive tolerance are blocked by intrathecal treatment with Raf-1-selective siRNA

    doi: 10.1111/j.1476-5381.2010.00869.x

    Figure Lengend Snippet: Intrathecal Raf-1-selective siRNA pretreatment attenuates sustained morphine-mediated tactile allodynia. For description of the animal groups see . Paw withdrawal thresholds in response to a series of von Frey filaments applied to the plantar surface

    Article Snippet: The rat Raf-1-selective siRNA mixture (Smart pool Dharmacon Inc., Chicago, IL, USA # L-087699-00) and the control, non-targeting double-stranded RNA (dsRNA) construct (Dharmacon Inc., Chicago, IL, USA #D-001810-01-20) were reconstituted in double distilled RNAse-free water to a stock concentration of 100 µM and stored in aliquots at −80°C.

    Techniques:

    Intrathecal (i.th) Raf-1-selective siRNA pretreatment attenuates sustained morphine-mediated antinociceptive tolerance. The groups of animals have been described in detail in . After sustained morphine (or saline) treatment, on day 6 the animals

    Journal: British Journal of Pharmacology

    Article Title: Sustained morphine-mediated pain sensitization and antinociceptive tolerance are blocked by intrathecal treatment with Raf-1-selective siRNA

    doi: 10.1111/j.1476-5381.2010.00869.x

    Figure Lengend Snippet: Intrathecal (i.th) Raf-1-selective siRNA pretreatment attenuates sustained morphine-mediated antinociceptive tolerance. The groups of animals have been described in detail in . After sustained morphine (or saline) treatment, on day 6 the animals

    Article Snippet: The rat Raf-1-selective siRNA mixture (Smart pool Dharmacon Inc., Chicago, IL, USA # L-087699-00) and the control, non-targeting double-stranded RNA (dsRNA) construct (Dharmacon Inc., Chicago, IL, USA #D-001810-01-20) were reconstituted in double distilled RNAse-free water to a stock concentration of 100 µM and stored in aliquots at −80°C.

    Techniques:

    Intrathecal (i.th) Raf-1-selective siRNA treatment attenuates sustained morphine-mediated augmentation of calcitonin gene-related peptide (CGRP) immunoreactivity in the lumbar spinal cord of rats. Male Sprague-Dawley rats received i.th vehicle (A, B)

    Journal: British Journal of Pharmacology

    Article Title: Sustained morphine-mediated pain sensitization and antinociceptive tolerance are blocked by intrathecal treatment with Raf-1-selective siRNA

    doi: 10.1111/j.1476-5381.2010.00869.x

    Figure Lengend Snippet: Intrathecal (i.th) Raf-1-selective siRNA treatment attenuates sustained morphine-mediated augmentation of calcitonin gene-related peptide (CGRP) immunoreactivity in the lumbar spinal cord of rats. Male Sprague-Dawley rats received i.th vehicle (A, B)

    Article Snippet: The rat Raf-1-selective siRNA mixture (Smart pool Dharmacon Inc., Chicago, IL, USA # L-087699-00) and the control, non-targeting double-stranded RNA (dsRNA) construct (Dharmacon Inc., Chicago, IL, USA #D-001810-01-20) were reconstituted in double distilled RNAse-free water to a stock concentration of 100 µM and stored in aliquots at −80°C.

    Techniques:

    Intrathecal Raf-1-selective siRNA treatment attenuates sustained morphine-mediated thermal hyperalgesia. The groups of rats included in the study were explained in . Paw withdrawal latencies in response to radiant heat applied to the plantar surface

    Journal: British Journal of Pharmacology

    Article Title: Sustained morphine-mediated pain sensitization and antinociceptive tolerance are blocked by intrathecal treatment with Raf-1-selective siRNA

    doi: 10.1111/j.1476-5381.2010.00869.x

    Figure Lengend Snippet: Intrathecal Raf-1-selective siRNA treatment attenuates sustained morphine-mediated thermal hyperalgesia. The groups of rats included in the study were explained in . Paw withdrawal latencies in response to radiant heat applied to the plantar surface

    Article Snippet: The rat Raf-1-selective siRNA mixture (Smart pool Dharmacon Inc., Chicago, IL, USA # L-087699-00) and the control, non-targeting double-stranded RNA (dsRNA) construct (Dharmacon Inc., Chicago, IL, USA #D-001810-01-20) were reconstituted in double distilled RNAse-free water to a stock concentration of 100 µM and stored in aliquots at −80°C.

    Techniques:

    Intrathecal Raf-1-selective siRNA pretreatment attenuates sustained morphine-mediated augmentation of calcitonin gene-related peptide (CGRP) levels in lumbar spinal cord homogenates of rats. Male Sprague-Dawley rats were subcutaneously implanted with

    Journal: British Journal of Pharmacology

    Article Title: Sustained morphine-mediated pain sensitization and antinociceptive tolerance are blocked by intrathecal treatment with Raf-1-selective siRNA

    doi: 10.1111/j.1476-5381.2010.00869.x

    Figure Lengend Snippet: Intrathecal Raf-1-selective siRNA pretreatment attenuates sustained morphine-mediated augmentation of calcitonin gene-related peptide (CGRP) levels in lumbar spinal cord homogenates of rats. Male Sprague-Dawley rats were subcutaneously implanted with

    Article Snippet: The rat Raf-1-selective siRNA mixture (Smart pool Dharmacon Inc., Chicago, IL, USA # L-087699-00) and the control, non-targeting double-stranded RNA (dsRNA) construct (Dharmacon Inc., Chicago, IL, USA #D-001810-01-20) were reconstituted in double distilled RNAse-free water to a stock concentration of 100 µM and stored in aliquots at −80°C.

    Techniques:

    (A) Intrathecal Raf-1-selective siRNA pretreatment reduces Raf-1 protein levels in rat lumbar spinal cord homogenates. Male Sprague-Dawley rats received intrathecal (i.th) injections of transfection reagent (lanes 1–2), 2 µg per 10 µL

    Journal: British Journal of Pharmacology

    Article Title: Sustained morphine-mediated pain sensitization and antinociceptive tolerance are blocked by intrathecal treatment with Raf-1-selective siRNA

    doi: 10.1111/j.1476-5381.2010.00869.x

    Figure Lengend Snippet: (A) Intrathecal Raf-1-selective siRNA pretreatment reduces Raf-1 protein levels in rat lumbar spinal cord homogenates. Male Sprague-Dawley rats received intrathecal (i.th) injections of transfection reagent (lanes 1–2), 2 µg per 10 µL

    Article Snippet: The rat Raf-1-selective siRNA mixture (Smart pool Dharmacon Inc., Chicago, IL, USA # L-087699-00) and the control, non-targeting double-stranded RNA (dsRNA) construct (Dharmacon Inc., Chicago, IL, USA #D-001810-01-20) were reconstituted in double distilled RNAse-free water to a stock concentration of 100 µM and stored in aliquots at −80°C.

    Techniques: Transfection

    A. Experimental design. (Ith: Intrathecal; siRNA: Raf-1 siRNA or mismatch siRNA delivery via intrathecal catheter; BL: Baseline; D: Day; IR: Infrared heat). B. Raf-1 siRNA attenuates sustained morphine-mediated thermal hyperalgesia in rats: Male Sprague Dawley rats received intrathecal injections of i-Fect encapsulated Raf-1selective siRNA (Raf-1 siRNA groups); or non-targeting dsRNA (mismatch siRNA groups) or the transfection agent (i-Fect) alone (control) once daily, for 3 days. After pre-treatment, the rats received continous subcutaneous (osmotic minipump) saline (control group, Raf-1 siRNA group and mismatch siRNA group) or morphine (45μg/μl/h) (morphine group, Raf-1 siRNA+morphine group and mismatch siRNA+morphine group) infusions for 6 days. Six animals were included in each treatment group. Thermal hyperalgesia was measured as a decrease in paw withdrawal latencies in a radiant heat paw-withdrawal test.

    Journal: European journal of pharmacology

    Article Title: Intrathecal Raf-1-selective siRNA attenuates sustained morphinemediated thermal hyperalgesia

    doi: 10.1016/j.ejphar.2008.10.033

    Figure Lengend Snippet: A. Experimental design. (Ith: Intrathecal; siRNA: Raf-1 siRNA or mismatch siRNA delivery via intrathecal catheter; BL: Baseline; D: Day; IR: Infrared heat). B. Raf-1 siRNA attenuates sustained morphine-mediated thermal hyperalgesia in rats: Male Sprague Dawley rats received intrathecal injections of i-Fect encapsulated Raf-1selective siRNA (Raf-1 siRNA groups); or non-targeting dsRNA (mismatch siRNA groups) or the transfection agent (i-Fect) alone (control) once daily, for 3 days. After pre-treatment, the rats received continous subcutaneous (osmotic minipump) saline (control group, Raf-1 siRNA group and mismatch siRNA group) or morphine (45μg/μl/h) (morphine group, Raf-1 siRNA+morphine group and mismatch siRNA+morphine group) infusions for 6 days. Six animals were included in each treatment group. Thermal hyperalgesia was measured as a decrease in paw withdrawal latencies in a radiant heat paw-withdrawal test.

    Article Snippet: After recovery from the surgery (5–7 days), the animals received intrathecal injections (2ug siRNA/10 ul/rat) of either a lipid encapsulated Raf-1-selective siRNA mixture (Smart pool siRNA, Dharmacon Inc; Chicago, IL, Cat # L-087699-00 ) (Raf-1 siRNA groups) or i-Fect encapsulated non-targeting dsRNA (Dharmacon, # D-001810-01-20 ) (control mismatch siRNA groups) or the transfection lipid alone, once daily, for 3 days, as described earlier ( ).

    Techniques: Transfection

    Raf-1 is essential for ERK activation by EGF and for PAK165-dependent rescue of ERK activation in cells treated with forskolin and IBMX. (A) Effects of Raf-1 siRNA. HEK293 cells were transfected with GFPERK, vector (−), or PAK165 (+) and

    Journal: Molecular and Cellular Biology

    Article Title: Raf-1 Serine 338 Phosphorylation Plays a Key Role in Adhesion-Dependent Activation of Extracellular Signal-Regulated Kinase by Epidermal Growth Factor †

    doi: 10.1128/MCB.25.11.4466-4475.2005

    Figure Lengend Snippet: Raf-1 is essential for ERK activation by EGF and for PAK165-dependent rescue of ERK activation in cells treated with forskolin and IBMX. (A) Effects of Raf-1 siRNA. HEK293 cells were transfected with GFPERK, vector (−), or PAK165 (+) and

    Article Snippet: Raf-1 small inhibitory RNA (siRNA) was obtained from Ambion (catalog number 51197; Huntingdon, United Kingdom).

    Techniques: Activation Assay, Transfection, Plasmid Preparation

    RRD-251 inhibits Rb-Raf-1 interaction in AoSMCs. (A and B) Treatment with TNFα or PDGF in the presence of RRD-251 inhibits Raf-1 levels in AoSMCs (A) and A10s (B). (C) TNFα and PDGF treatment induced Rb-Raf-1 binding in AoSMCs. (D) RRD-251 inhibits TNFα-induced Rb-Raf-1 interaction.

    Journal: Cell Cycle

    Article Title: TNF?-mediated proliferation of vascular smooth muscle cells involves Raf-1-mediated inactivation of Rb and transcription of E2F1-regulated genes

    doi: 10.4161/cc.11.1.18473

    Figure Lengend Snippet: RRD-251 inhibits Rb-Raf-1 interaction in AoSMCs. (A and B) Treatment with TNFα or PDGF in the presence of RRD-251 inhibits Raf-1 levels in AoSMCs (A) and A10s (B). (C) TNFα and PDGF treatment induced Rb-Raf-1 binding in AoSMCs. (D) RRD-251 inhibits TNFα-induced Rb-Raf-1 interaction.

    Article Snippet: siRNA oligonucleotides targeting human E2F1, E2F2, E2F3 and Raf-1 were purchased from Santa Cruz biotechnology.

    Techniques: Binding Assay

    Whereas in AoSMCs, TNFα stimulation led to Raf-1-dependent Rb inactivation, elevated E2F1 activity, leading to the expression of proliferative promoters, resulting in increased cell proliferation.

    Journal: Cell Cycle

    Article Title: TNF?-mediated proliferation of vascular smooth muscle cells involves Raf-1-mediated inactivation of Rb and transcription of E2F1-regulated genes

    doi: 10.4161/cc.11.1.18473

    Figure Lengend Snippet: Whereas in AoSMCs, TNFα stimulation led to Raf-1-dependent Rb inactivation, elevated E2F1 activity, leading to the expression of proliferative promoters, resulting in increased cell proliferation.

    Article Snippet: siRNA oligonucleotides targeting human E2F1, E2F2, E2F3 and Raf-1 were purchased from Santa Cruz biotechnology.

    Techniques: Activity Assay, Expressing

    TNFα activates the Raf/MAPK pathway in VSMCs. (A) Time-course stimulation of AoSMCs results in Raf-1 activation, highest at 1 h, and ERK1/2 activation peaks at 30 min. (B) Activation of ERK1/2 coincides with JNK1 activation from 30 min of TNFα treatment. (C) PDGF and TNFα time-course stimulation shows ERK1/2 and JNK activation occurs simultaneously from the different stimuli.

    Journal: Cell Cycle

    Article Title: TNF?-mediated proliferation of vascular smooth muscle cells involves Raf-1-mediated inactivation of Rb and transcription of E2F1-regulated genes

    doi: 10.4161/cc.11.1.18473

    Figure Lengend Snippet: TNFα activates the Raf/MAPK pathway in VSMCs. (A) Time-course stimulation of AoSMCs results in Raf-1 activation, highest at 1 h, and ERK1/2 activation peaks at 30 min. (B) Activation of ERK1/2 coincides with JNK1 activation from 30 min of TNFα treatment. (C) PDGF and TNFα time-course stimulation shows ERK1/2 and JNK activation occurs simultaneously from the different stimuli.

    Article Snippet: siRNA oligonucleotides targeting human E2F1, E2F2, E2F3 and Raf-1 were purchased from Santa Cruz biotechnology.

    Techniques: Activation Assay

    Targeting Raf-1 activation blocks AoSMC proliferation. (A) Src inhibitor (PP2) and PKC inhibitor (Ro-31-8220) block TNFα- and PDGF-induced proliferation. (B) Multi-kinase inhibitor BAY-43-9006, which targets Raf-1, can completely inhibit PDGF- and TNFα-induced proliferation, while the JNK inhibitor (SP600125) does not significantly affect TNFα-induced proliferation. (C) MEK inhibitor PD98059 could block TNFα- and PDGF-induced S-phase entry as seen by BrdU incorporation assays. (D) Raf-1 depletion using Raf-1 siRNAs could inhibit TNFα- and PDGF-induced AoSMC proliferation.

    Journal: Cell Cycle

    Article Title: TNF?-mediated proliferation of vascular smooth muscle cells involves Raf-1-mediated inactivation of Rb and transcription of E2F1-regulated genes

    doi: 10.4161/cc.11.1.18473

    Figure Lengend Snippet: Targeting Raf-1 activation blocks AoSMC proliferation. (A) Src inhibitor (PP2) and PKC inhibitor (Ro-31-8220) block TNFα- and PDGF-induced proliferation. (B) Multi-kinase inhibitor BAY-43-9006, which targets Raf-1, can completely inhibit PDGF- and TNFα-induced proliferation, while the JNK inhibitor (SP600125) does not significantly affect TNFα-induced proliferation. (C) MEK inhibitor PD98059 could block TNFα- and PDGF-induced S-phase entry as seen by BrdU incorporation assays. (D) Raf-1 depletion using Raf-1 siRNAs could inhibit TNFα- and PDGF-induced AoSMC proliferation.

    Article Snippet: siRNA oligonucleotides targeting human E2F1, E2F2, E2F3 and Raf-1 were purchased from Santa Cruz biotechnology.

    Techniques: Activation Assay, Blocking Assay, BrdU Incorporation Assay

    TNFα and PDGF induce E2F regulated genes in AoSMCs. (A) Treatment with TNFα and PDGF for 18 h led to 3.5- and 4-fold increase, respectively, in cdc25A gene expression in real-time PCR assays. (B) Treatment with TNFα and PDGF for 18 h led to 3.5- and 7-fold increase, respectively in cdc6 gene expression in real-time PCR assays. (C) Treatment with TNFα and PDGF for 18 h led to 1.8 and 3.8 fold increase, respectively in TS gene expression in real time PCR assays. (D) Treatment with TNFα or PDGF led to an increase in E2F1 and dissociation of Rb on the proliferative promoters cdc25A, cdc6 and TS in ChIP assays; c-fos was used as the negative control. (E) Western blots for cdc 6, cdc 25A and TS after Raf-1 and E2F1 depletion. (F) Densitometric analysis of the bands to quantify the western blot data.

    Journal: Cell Cycle

    Article Title: TNF?-mediated proliferation of vascular smooth muscle cells involves Raf-1-mediated inactivation of Rb and transcription of E2F1-regulated genes

    doi: 10.4161/cc.11.1.18473

    Figure Lengend Snippet: TNFα and PDGF induce E2F regulated genes in AoSMCs. (A) Treatment with TNFα and PDGF for 18 h led to 3.5- and 4-fold increase, respectively, in cdc25A gene expression in real-time PCR assays. (B) Treatment with TNFα and PDGF for 18 h led to 3.5- and 7-fold increase, respectively in cdc6 gene expression in real-time PCR assays. (C) Treatment with TNFα and PDGF for 18 h led to 1.8 and 3.8 fold increase, respectively in TS gene expression in real time PCR assays. (D) Treatment with TNFα or PDGF led to an increase in E2F1 and dissociation of Rb on the proliferative promoters cdc25A, cdc6 and TS in ChIP assays; c-fos was used as the negative control. (E) Western blots for cdc 6, cdc 25A and TS after Raf-1 and E2F1 depletion. (F) Densitometric analysis of the bands to quantify the western blot data.

    Article Snippet: siRNA oligonucleotides targeting human E2F1, E2F2, E2F3 and Raf-1 were purchased from Santa Cruz biotechnology.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation, Negative Control, Western Blot