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  • 99
    Thermo Fisher rt pcr
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    Qiagen onestep rt pcr kit
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    Bio-Rad real time reverse transcription polymerase chain reaction
    Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a <t>time</t> point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using <t>real-time</t> <t>polymerase</t> <t>chain</t> <t>reaction</t> in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P
    Real Time Reverse Transcription Polymerase Chain Reaction, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen quantitect sybr green rt pcr kit
    Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a <t>time</t> point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using <t>real-time</t> <t>polymerase</t> <t>chain</t> <t>reaction</t> in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P
    Quantitect Sybr Green Rt Pcr Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 8359 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad quantitative reverse transcription polymerase chain reaction
    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
    Quantitative Reverse Transcription Polymerase Chain Reaction, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rt pcr total rna
    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
    Rt Pcr Total Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 12883 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher rt pcr kit
    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
    Rt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6074 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa primescript rt pcr kit
    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
    Primescript Rt Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 4795 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad iscript one step rt pcr kit
    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
    Iscript One Step Rt Pcr Kit, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 5922 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
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    Qiagen quantitect probe rt pcr kit
    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
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    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
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    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
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    Stratagene rt pcr
    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
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    Qiagen quantifast sybr green rt pcr kit
    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
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    TaKaRa sybr primescript rt pcr kit
    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. <t>Reverse</t> <t>transcription–polymerase</t> <t>chain</t> <t>reaction</t> showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.
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    Image Search Results


    Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P

    Journal: Carcinogenesis

    Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

    doi: 10.1093/carcin/bgr075

    Figure Lengend Snippet: Auto/paracrine IL-6 trans-signaling is enhanced in MIA-MSLN cells through upregulated soluble IL-6 receptors. ( A ) Exogenous rIL-6 induces proliferation in MIA-MSLN cells. MIA-MSLN and control MIA-V cells were seeded in 96-well plates (2 × 10 3 cells per well), serum starved (0% FBS) for 24 h and treated with indicated concentrations of IL-6 in 0.2% FBS containing medium for 3 days. Viability was measured with MTT. Relative increase in viability was measured by dividing viability at a time point by viability of same cells at day 0 (day after plating) and is plotted along Y -axis. Data plotted show mean of triplicate wells and is representative of at least two similar experiments. ( B ) IL-6 receptor gp80 and gp130 expression patterns in MIA-V and MIA-MSLN cells. IL-6 receptor gp80 and gp130 mRNA expression levels were determined by using real-time polymerase chain reaction in both MIA-V and MIA-MSLN cells. Y -axis shows GAPDH-normalized mRNA levels for gp80 and gp130 in MIA-V and MIA-MSLN cells. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. ( C ) Cell surface IL-6R (gp80) and soluble IL-6 receptor (sIL-6R) expression patterns in MIA-V and MIA-MSLN cells. (a) Cell surface gp80 expression was analyzed by FACS using anti-gp80 antibody. The results are depicted as histograms, the percentage denotes the positive cell population. sIL-6R expression in MIA-V and MIA-MSLN supernatants (b) was analyzed by western blot with anti-IL-6R antibody. *, # denote P

    Article Snippet: MSLN, IL-6, IL-6R and sIL-6R (alternatively spliced form), ADAM10, ADAM17 and GAPDH messenger RNA (mRNAs) were analyzed by real-time reverse transcription–polymerase chain reaction using the SYBR Green supermix kit (Bio-Rad Laboratories) as described previously ( ).

    Techniques: MTT Assay, Expressing, Real-time Polymerase Chain Reaction, FACS, Western Blot

    MSLN overexpression leads to increased IL-6 production in PC cells. ( A ). MIA, MIA-V, MIA-GFP and MIA-MSLN cells were seeded at 1 × 10 6 cells per well in six-well plates and cultured until 60% confluence. Thereafter, the medium was replaced and the supernatants were harvested at 48 h of further incubation and the levels of IL-6 were determined by using the Luminex-based IL-6 assay kit. Y -axis represents IL-6 concentration in pg/ml. ( B ) Silencing MSLN using MSLN-specific shRNA plasmid. MIA-V and MIA-MSLN cells were transfected with MSLN-specific shRNA plasmids or GFP shRNA control plasmids. The cells collected 48 h posttransfection were used to detect MSLN mRNA by real-time polymerase chain reaction. The MSLN-expressing levels were detected by using real-time polymerase chain reaction. Y -axis represents MSLN mRNA level normalized to the GAPDH mRNA level. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. Experiment was performed several times using separate MIA-MSLN pools and different siRNA/shRNAs against MSLN with similar results. ( C ) Silencing MSLN decreases IL-6 mRNA production in MIA-MSLN cells. MIA-V and MIA-MSLN cells were transfected with MSLN-specific shRNA plasmids or GFP shRNA control plasmids. The cells collected 48 h posttransfection were used to detect IL-6 mRNA by real-time polymerase chain reaction. Y -axis represents IL-6 mRNA level relative to GAPDH. The bars denote SD of duplicate data. Experiment was performed several times using separate MIA-MSLN pools and different siRNA/shRNAs against MSLN with similar results. ( D ) Silencing MSLN decreases IL-6 secretion in MIA-MSLN cells. MIA-V and MIA-MSLN cells were transfected with MSLN-specific siRNA or scrambled siRNA. The cells collected 48 h posttransfection from 24-well plates in 2 ml of medium were used to detect IL-6 by Luminex-based IL-6 assay kit. Values on Y -axis show the amount of IL-6 in pg/ml, bars denoting SD of duplicate data. ( E ) Stable overexpression of MSLN in human PC cell Panc1. The GAPDH-normalized MSLN expression levels in the stably MSLN expressing (Panc1-MSLN) and control MIA cells generated by retroviral gene transfer and subsequent puromycin selection are shown. ( F ) Panc1, Panc1-V, Panc1-GFP and Panc1-MSLN cells were seeded at 1 × 10 6 cells per well in six-well plates and cultured until 80% confluence, cells collected were used to detect IL-6 mRNA by real-time polymerase chain reaction. Y -axis represents IL-6 mRNA level relative to GAPDH. The bars denote SD of duplicate data. *, # denote P

    Journal: Carcinogenesis

    Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

    doi: 10.1093/carcin/bgr075

    Figure Lengend Snippet: MSLN overexpression leads to increased IL-6 production in PC cells. ( A ). MIA, MIA-V, MIA-GFP and MIA-MSLN cells were seeded at 1 × 10 6 cells per well in six-well plates and cultured until 60% confluence. Thereafter, the medium was replaced and the supernatants were harvested at 48 h of further incubation and the levels of IL-6 were determined by using the Luminex-based IL-6 assay kit. Y -axis represents IL-6 concentration in pg/ml. ( B ) Silencing MSLN using MSLN-specific shRNA plasmid. MIA-V and MIA-MSLN cells were transfected with MSLN-specific shRNA plasmids or GFP shRNA control plasmids. The cells collected 48 h posttransfection were used to detect MSLN mRNA by real-time polymerase chain reaction. The MSLN-expressing levels were detected by using real-time polymerase chain reaction. Y -axis represents MSLN mRNA level normalized to the GAPDH mRNA level. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (IL-6)]. The bars denote SD of duplicate data. Experiment was performed several times using separate MIA-MSLN pools and different siRNA/shRNAs against MSLN with similar results. ( C ) Silencing MSLN decreases IL-6 mRNA production in MIA-MSLN cells. MIA-V and MIA-MSLN cells were transfected with MSLN-specific shRNA plasmids or GFP shRNA control plasmids. The cells collected 48 h posttransfection were used to detect IL-6 mRNA by real-time polymerase chain reaction. Y -axis represents IL-6 mRNA level relative to GAPDH. The bars denote SD of duplicate data. Experiment was performed several times using separate MIA-MSLN pools and different siRNA/shRNAs against MSLN with similar results. ( D ) Silencing MSLN decreases IL-6 secretion in MIA-MSLN cells. MIA-V and MIA-MSLN cells were transfected with MSLN-specific siRNA or scrambled siRNA. The cells collected 48 h posttransfection from 24-well plates in 2 ml of medium were used to detect IL-6 by Luminex-based IL-6 assay kit. Values on Y -axis show the amount of IL-6 in pg/ml, bars denoting SD of duplicate data. ( E ) Stable overexpression of MSLN in human PC cell Panc1. The GAPDH-normalized MSLN expression levels in the stably MSLN expressing (Panc1-MSLN) and control MIA cells generated by retroviral gene transfer and subsequent puromycin selection are shown. ( F ) Panc1, Panc1-V, Panc1-GFP and Panc1-MSLN cells were seeded at 1 × 10 6 cells per well in six-well plates and cultured until 80% confluence, cells collected were used to detect IL-6 mRNA by real-time polymerase chain reaction. Y -axis represents IL-6 mRNA level relative to GAPDH. The bars denote SD of duplicate data. *, # denote P

    Article Snippet: MSLN, IL-6, IL-6R and sIL-6R (alternatively spliced form), ADAM10, ADAM17 and GAPDH messenger RNA (mRNAs) were analyzed by real-time reverse transcription–polymerase chain reaction using the SYBR Green supermix kit (Bio-Rad Laboratories) as described previously ( ).

    Techniques: Over Expression, Cell Culture, Incubation, Luminex, Concentration Assay, shRNA, Plasmid Preparation, Transfection, Real-time Polymerase Chain Reaction, Expressing, Stable Transfection, Generated, Selection

    MSLN expression level correlates well with IL-6 expression in PC cell lines and tissues. ( A ) IL-6 levels in PC cell lines MIA PaCa-2, Panc-1, BxPC-3 and HPDE cells. Cells were seeded at 1 × 10 6 cells per well in six-well plates and cultured until 60% confluence. Thereafter, the medium was replaced and the supernatants were harvested at 48 h of further incubation and tested for IL-6 using the Luminex-based IL-6 assay kit. Values on Y -axis show the amount of IL-6 in pg/ml, bars denoting standard deviation (SD) of duplicate data. ( B ) Relative MSLN mRNA levels in PC cell lines MIA PaCa-2, Panc-1, BxPC-3 and HPDE cells. Total mRNA from the cell lines were reverse transcribed and tested for MSLN expression. The results depicted denote MSLN mRNA levels in each cell line normalized to the GAPDH mRNA level. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (MSLN)] and is representative of at least two independent experiments. The bars denote SD of duplicate data. ( C ) Sera from six PC patients were tested for secreted IL-6 using Bioplex cytokine kit. Paired tumor and normal tissues (T/N) from the same patients were tested for the expression of MSLN mRNA by using real-time polymerase chain reaction and represented by the bar graph. The left hand side Y -axis denotes T/N ratio of the GAPDH-normalized MSLN mRNA levels. Line graph represents the serum IL-6 level in six patient samples and the right hand side Y -axis indicates IL-6 concentrations in pg/ml.

    Journal: Carcinogenesis

    Article Title: Mesothelin overexpression promotes autocrine IL-6/sIL-6R trans-signaling to stimulate pancreatic cancer cell proliferation

    doi: 10.1093/carcin/bgr075

    Figure Lengend Snippet: MSLN expression level correlates well with IL-6 expression in PC cell lines and tissues. ( A ) IL-6 levels in PC cell lines MIA PaCa-2, Panc-1, BxPC-3 and HPDE cells. Cells were seeded at 1 × 10 6 cells per well in six-well plates and cultured until 60% confluence. Thereafter, the medium was replaced and the supernatants were harvested at 48 h of further incubation and tested for IL-6 using the Luminex-based IL-6 assay kit. Values on Y -axis show the amount of IL-6 in pg/ml, bars denoting standard deviation (SD) of duplicate data. ( B ) Relative MSLN mRNA levels in PC cell lines MIA PaCa-2, Panc-1, BxPC-3 and HPDE cells. Total mRNA from the cell lines were reverse transcribed and tested for MSLN expression. The results depicted denote MSLN mRNA levels in each cell line normalized to the GAPDH mRNA level. Relative mRNA level is presented as 2 [ C t (GAPDH)− C t (MSLN)] and is representative of at least two independent experiments. The bars denote SD of duplicate data. ( C ) Sera from six PC patients were tested for secreted IL-6 using Bioplex cytokine kit. Paired tumor and normal tissues (T/N) from the same patients were tested for the expression of MSLN mRNA by using real-time polymerase chain reaction and represented by the bar graph. The left hand side Y -axis denotes T/N ratio of the GAPDH-normalized MSLN mRNA levels. Line graph represents the serum IL-6 level in six patient samples and the right hand side Y -axis indicates IL-6 concentrations in pg/ml.

    Article Snippet: MSLN, IL-6, IL-6R and sIL-6R (alternatively spliced form), ADAM10, ADAM17 and GAPDH messenger RNA (mRNAs) were analyzed by real-time reverse transcription–polymerase chain reaction using the SYBR Green supermix kit (Bio-Rad Laboratories) as described previously ( ).

    Techniques: Expressing, Cell Culture, Incubation, Luminex, Standard Deviation, Real-time Polymerase Chain Reaction

    SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. Reverse transcription–polymerase chain reaction showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.

    Journal: Carcinogenesis

    Article Title: Use of a TGF? type I receptor inhibitor in mouse skin carcinogenesis reveals a dual role for TGF? signaling in tumor promotion and progression

    doi: 10.1093/carcin/bgq191

    Figure Lengend Snippet: SB suppresses TPA-induced inflammation. ( A ) Effect of SB on chronic TPA-induced hyperplasia (top) proliferation (middle) and apoptosis (bottom) ( B ) Decreased cutaneous neutrophils (MPO+ cells) after acute and chronic TPA + SB treatment. ( C ) Decreased keratinocyte chemokine (KC) and macrophage-inflammatory protein 2 (MIP2) gene expression in acute and chronically SB- and TPA-treated whole skin. Results normalized to TPA only samples. Reverse transcription–polymerase chain reaction showing induction of these chemokines by TPA (bottom). n = 4–6 mice per treatment group. A, acetone.

    Article Snippet: Total RNA was isolated from whole skin using Trizol (Invitrogen, Carlsbad, CA) and quantitative reverse transcription–polymerase chain reaction done for the indicated genes using the MyIQ system (Bio-Rad Laboratories, Hercules, CA).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Mouse Assay

    SB rapidly induces progressed phenotype in epidermis-expressing human C-Ha RASV12G oncogene. ( A ) Significant increase in proliferation with SB treatment in RAS -expressing skin (left) and slight increase in epidermal thickness (right). ( B ) An increase in MPO+ cells in the dermis of SB-treated mice expressing the InvtTA × tetO RASV12G transgenes. Arrows indicate neutrophils. Yellow line shows epidermal/dermal junction. Representative micrographs (right), magnification, ×100. Scale bar represents 50 μm. ( C ) Increased messenger RNA expression of neutrophil chemokines s100a8, s100a9 and keratinocyte chemokine with treatment of SB. ( D ) SB causes significant decrease in keratin 1 staining (left) and messenger RNA expression (middle). Representative micrographs (left), magnification ×100. Scale bar represents 50 μm. Yellow dotted line shows epidermal/dermal junction. Significant increase in keratin 18 messenger RNA expression by quantitative reverse transcription–polymerase chain reaction in RAS -expressing skin following SB (right). n = 4–6 mice per treatment group.

    Journal: Carcinogenesis

    Article Title: Use of a TGF? type I receptor inhibitor in mouse skin carcinogenesis reveals a dual role for TGF? signaling in tumor promotion and progression

    doi: 10.1093/carcin/bgq191

    Figure Lengend Snippet: SB rapidly induces progressed phenotype in epidermis-expressing human C-Ha RASV12G oncogene. ( A ) Significant increase in proliferation with SB treatment in RAS -expressing skin (left) and slight increase in epidermal thickness (right). ( B ) An increase in MPO+ cells in the dermis of SB-treated mice expressing the InvtTA × tetO RASV12G transgenes. Arrows indicate neutrophils. Yellow line shows epidermal/dermal junction. Representative micrographs (right), magnification, ×100. Scale bar represents 50 μm. ( C ) Increased messenger RNA expression of neutrophil chemokines s100a8, s100a9 and keratinocyte chemokine with treatment of SB. ( D ) SB causes significant decrease in keratin 1 staining (left) and messenger RNA expression (middle). Representative micrographs (left), magnification ×100. Scale bar represents 50 μm. Yellow dotted line shows epidermal/dermal junction. Significant increase in keratin 18 messenger RNA expression by quantitative reverse transcription–polymerase chain reaction in RAS -expressing skin following SB (right). n = 4–6 mice per treatment group.

    Article Snippet: Total RNA was isolated from whole skin using Trizol (Invitrogen, Carlsbad, CA) and quantitative reverse transcription–polymerase chain reaction done for the indicated genes using the MyIQ system (Bio-Rad Laboratories, Hercules, CA).

    Techniques: Expressing, Mouse Assay, RNA Expression, Staining, Reverse Transcription Polymerase Chain Reaction

    SB reduces TPA-induced Smad2 phosphorylation in vivo . ( A (available at Carcinogenesis Online). ( B ) Quantitative reverse transcription–polymerase chain reaction analysis of Smad7 and LTBP-1 gene expression in acetone or SB-treated whole skin. Normalized to acetone only samples ( n = 4). ( C ) Indirect immunofluorescence showing a reduction in TPA-induced p-Smad2 by SB at 4 and 24 h TPA treatment. Magnification ×800, bar represents 15 μm. P-Smad2 was detected with Alexa-fluor-488 (green), and nuclei counterstained with TO-PRO3 (red), (representative of n (available at Carcinogenesis Online). ( D ) Western blot of whole skin treated 24 h with TPA and 12 h SB, showing reduced TPA-induced Smad2 phosphorylation with SB treatment (representative of n (available at Carcinogenesis Online). C, control and A, acetone.

    Journal: Carcinogenesis

    Article Title: Use of a TGF? type I receptor inhibitor in mouse skin carcinogenesis reveals a dual role for TGF? signaling in tumor promotion and progression

    doi: 10.1093/carcin/bgq191

    Figure Lengend Snippet: SB reduces TPA-induced Smad2 phosphorylation in vivo . ( A (available at Carcinogenesis Online). ( B ) Quantitative reverse transcription–polymerase chain reaction analysis of Smad7 and LTBP-1 gene expression in acetone or SB-treated whole skin. Normalized to acetone only samples ( n = 4). ( C ) Indirect immunofluorescence showing a reduction in TPA-induced p-Smad2 by SB at 4 and 24 h TPA treatment. Magnification ×800, bar represents 15 μm. P-Smad2 was detected with Alexa-fluor-488 (green), and nuclei counterstained with TO-PRO3 (red), (representative of n (available at Carcinogenesis Online). ( D ) Western blot of whole skin treated 24 h with TPA and 12 h SB, showing reduced TPA-induced Smad2 phosphorylation with SB treatment (representative of n (available at Carcinogenesis Online). C, control and A, acetone.

    Article Snippet: Total RNA was isolated from whole skin using Trizol (Invitrogen, Carlsbad, CA) and quantitative reverse transcription–polymerase chain reaction done for the indicated genes using the MyIQ system (Bio-Rad Laboratories, Hercules, CA).

    Techniques: In Vivo, Reverse Transcription Polymerase Chain Reaction, Expressing, Immunofluorescence, Western Blot