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    Thermo Fisher real time pcr system
    Validation of vorinostat-induced transcriptional responses by <t>qRT-PCR.</t> Transcriptional responses of BJ and BJ LTSTERas fibroblasts following DMSO (black) and 25 μ M vorinostat treatment (red) were validated by qRT-PCR using the same template RNA as that used in the microarray study. Log2 transformed expression intensities of HG U133 plus 2.0 probe sets corresponding to ( a ) BAK1 , BAD, BIK and BCL2A1 and ( b ) BCL2L11 and BMF are shown. Gene names and probe set IDs are indicated in the title of each panel. Normalized probe set intensity values ( Y -axis) are presented, which allows direct comparison of basal expression (time 0 h), and log2 transcriptional responses over time between cell types (BJ and BJ LTSTERas). Data represent the summarized probe set intensities from three independent microarray time course experiments. qRT-PCR verification of selected genes is shown. Transcript expression was calculated relative to the control gene RPL32 (that does not respond to vorinostat or DMSO treatment), and fold-change mRNA expression over time was calculated relative to the 0 h time point. Data are presented as the mean±S.E.M. of three independent experiments. For both Affymetrix and qRT-PCR results, data sets for BJ cells are in the left panels while data sets for BJ LTSTERas are in the right panels. ( c ) Whole cell protein extracts from BJ and BJ LTSTERas fibroblasts treated with DMSO or 25 μ M vorinostat for 0–48 h were separated by <t>SDS-PAGE</t> and analyzed for the expression of BMF, BIK and BIM by western blotting. The expression of α -Tubulin was analyzed to confirm equal protein loading in each lane. The asterisk (*) denotes a non-specific band in the BIK western blot, which acts as a positive control for activity of the BIK antibody. The arrow denotes the location of BIK. Images are representative of two independent experiments
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    Images

    1) Product Images from "HDAC inhibitors induce tumor-cell-selective pro-apoptotic transcriptional responses"

    Article Title: HDAC inhibitors induce tumor-cell-selective pro-apoptotic transcriptional responses

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2013.9

    Validation of vorinostat-induced transcriptional responses by qRT-PCR. Transcriptional responses of BJ and BJ LTSTERas fibroblasts following DMSO (black) and 25 μ M vorinostat treatment (red) were validated by qRT-PCR using the same template RNA as that used in the microarray study. Log2 transformed expression intensities of HG U133 plus 2.0 probe sets corresponding to ( a ) BAK1 , BAD, BIK and BCL2A1 and ( b ) BCL2L11 and BMF are shown. Gene names and probe set IDs are indicated in the title of each panel. Normalized probe set intensity values ( Y -axis) are presented, which allows direct comparison of basal expression (time 0 h), and log2 transcriptional responses over time between cell types (BJ and BJ LTSTERas). Data represent the summarized probe set intensities from three independent microarray time course experiments. qRT-PCR verification of selected genes is shown. Transcript expression was calculated relative to the control gene RPL32 (that does not respond to vorinostat or DMSO treatment), and fold-change mRNA expression over time was calculated relative to the 0 h time point. Data are presented as the mean±S.E.M. of three independent experiments. For both Affymetrix and qRT-PCR results, data sets for BJ cells are in the left panels while data sets for BJ LTSTERas are in the right panels. ( c ) Whole cell protein extracts from BJ and BJ LTSTERas fibroblasts treated with DMSO or 25 μ M vorinostat for 0–48 h were separated by SDS-PAGE and analyzed for the expression of BMF, BIK and BIM by western blotting. The expression of α -Tubulin was analyzed to confirm equal protein loading in each lane. The asterisk (*) denotes a non-specific band in the BIK western blot, which acts as a positive control for activity of the BIK antibody. The arrow denotes the location of BIK. Images are representative of two independent experiments
    Figure Legend Snippet: Validation of vorinostat-induced transcriptional responses by qRT-PCR. Transcriptional responses of BJ and BJ LTSTERas fibroblasts following DMSO (black) and 25 μ M vorinostat treatment (red) were validated by qRT-PCR using the same template RNA as that used in the microarray study. Log2 transformed expression intensities of HG U133 plus 2.0 probe sets corresponding to ( a ) BAK1 , BAD, BIK and BCL2A1 and ( b ) BCL2L11 and BMF are shown. Gene names and probe set IDs are indicated in the title of each panel. Normalized probe set intensity values ( Y -axis) are presented, which allows direct comparison of basal expression (time 0 h), and log2 transcriptional responses over time between cell types (BJ and BJ LTSTERas). Data represent the summarized probe set intensities from three independent microarray time course experiments. qRT-PCR verification of selected genes is shown. Transcript expression was calculated relative to the control gene RPL32 (that does not respond to vorinostat or DMSO treatment), and fold-change mRNA expression over time was calculated relative to the 0 h time point. Data are presented as the mean±S.E.M. of three independent experiments. For both Affymetrix and qRT-PCR results, data sets for BJ cells are in the left panels while data sets for BJ LTSTERas are in the right panels. ( c ) Whole cell protein extracts from BJ and BJ LTSTERas fibroblasts treated with DMSO or 25 μ M vorinostat for 0–48 h were separated by SDS-PAGE and analyzed for the expression of BMF, BIK and BIM by western blotting. The expression of α -Tubulin was analyzed to confirm equal protein loading in each lane. The asterisk (*) denotes a non-specific band in the BIK western blot, which acts as a positive control for activity of the BIK antibody. The arrow denotes the location of BIK. Images are representative of two independent experiments

    Techniques Used: Quantitative RT-PCR, Microarray, Transformation Assay, Expressing, SDS Page, Western Blot, Positive Control, Activity Assay

    2) Product Images from "Rapid enhancement of α-tocopherol content in Nicotiana benthamiana by transient expression of Arabidopsis thaliana Tocopherol cyclase and Homogentisate phytyl transferase genes"

    Article Title: Rapid enhancement of α-tocopherol content in Nicotiana benthamiana by transient expression of Arabidopsis thaliana Tocopherol cyclase and Homogentisate phytyl transferase genes

    Journal: 3 Biotech

    doi: 10.1007/s13205-018-1496-4

    Transient expression of AtHPT, AtTC and AtTC + AtHPT post agroinfiltration by reverse transcription PCR. Top panel: Target genes Lane 1 Wild type control Lane 2 HPT Lane 3 TC Lane 3 HPT + TC with TC primers Lane 3 HPT + TC with HPT primers Lane 6 HPT + TC with both HPT TC primers. Bottom panel: Reference gene F-box
    Figure Legend Snippet: Transient expression of AtHPT, AtTC and AtTC + AtHPT post agroinfiltration by reverse transcription PCR. Top panel: Target genes Lane 1 Wild type control Lane 2 HPT Lane 3 TC Lane 3 HPT + TC with TC primers Lane 3 HPT + TC with HPT primers Lane 6 HPT + TC with both HPT TC primers. Bottom panel: Reference gene F-box

    Techniques Used: Expressing, Polymerase Chain Reaction

    3) Product Images from "HIF-2α mediates hypoxia-induced LIF expression in human colorectal cancer cells"

    Article Title: HIF-2α mediates hypoxia-induced LIF expression in human colorectal cancer cells

    Journal: Oncotarget

    doi:

    Hypoxia transactivates hypoxia-responsive elements (HREs) in the LIF promoter through HIF-2α (A) The human LIF gene contains 4 putative HREs in its promoter region. (B) Hypoxia activates the luciferase activity of reporter vectors containing HRE-C or HRE-D sites in the LIF promoter. RKO and HCT116 cells were transfected with the luciferase reporter vectors, and then subjected to hypoxia treatment for 36 h before measuring luciferase activities. Luciferase reporter vectors containing the HRE site in the VEGF promoter was included as a positive control. (C) and (D) HIF-2α but not HIF-1α binds to HRE-C and HRE-D sites in the LIF promoter under the hypoxic condition in RKO cells as determined by ChIP assays. Cells were cultured under the hypoxic or normoxic conditions for 36 h before assays. The HRE site in the VEGF promoter serves as a positive control. The amount of DNA fragments pulled-down was determined by real-time PCR (C) or conventional PCR (D) . Data are presented as mean ± SD ( n = 3). *: p
    Figure Legend Snippet: Hypoxia transactivates hypoxia-responsive elements (HREs) in the LIF promoter through HIF-2α (A) The human LIF gene contains 4 putative HREs in its promoter region. (B) Hypoxia activates the luciferase activity of reporter vectors containing HRE-C or HRE-D sites in the LIF promoter. RKO and HCT116 cells were transfected with the luciferase reporter vectors, and then subjected to hypoxia treatment for 36 h before measuring luciferase activities. Luciferase reporter vectors containing the HRE site in the VEGF promoter was included as a positive control. (C) and (D) HIF-2α but not HIF-1α binds to HRE-C and HRE-D sites in the LIF promoter under the hypoxic condition in RKO cells as determined by ChIP assays. Cells were cultured under the hypoxic or normoxic conditions for 36 h before assays. The HRE site in the VEGF promoter serves as a positive control. The amount of DNA fragments pulled-down was determined by real-time PCR (C) or conventional PCR (D) . Data are presented as mean ± SD ( n = 3). *: p

    Techniques Used: Luciferase, Activity Assay, Transfection, Positive Control, Chromatin Immunoprecipitation, Cell Culture, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction

    Hypoxia induces LIF expression levels in human colorectal cancer cell lines Human colorectal cancer cell lines RKO and HCT116 cells were cultured under the hypoxic condition for the indicated time periods. (A) The mRNA expression levels of LIF in these cells were determined by Taqman real-time PCR and normalized with actin (left panels). The mRNA expression levels of VEGF in these cells were determined as a positive control (right panels). (B) The LIF protein levels were determined by Elisa assays (left panels) and Western-blot assays (right panels), respectively. Data are presented as mean ± SD ( n = 3). *: p
    Figure Legend Snippet: Hypoxia induces LIF expression levels in human colorectal cancer cell lines Human colorectal cancer cell lines RKO and HCT116 cells were cultured under the hypoxic condition for the indicated time periods. (A) The mRNA expression levels of LIF in these cells were determined by Taqman real-time PCR and normalized with actin (left panels). The mRNA expression levels of VEGF in these cells were determined as a positive control (right panels). (B) The LIF protein levels were determined by Elisa assays (left panels) and Western-blot assays (right panels), respectively. Data are presented as mean ± SD ( n = 3). *: p

    Techniques Used: Expressing, Cell Culture, Real-time Polymerase Chain Reaction, Positive Control, Enzyme-linked Immunosorbent Assay, Western Blot

    HIF-2α transcriptionally regulates LIF expression (A) Ectopic HIF-2α expression increases LIF and VEGF mRNA expression levels in RKO and HCT116 cells. The mRNA expression levels of LIF and VEGF were determined by Taqman real-time PCR and normalized with actin. (B) Ectopic HIF-1α expression increases VEGF mRNA expression levels but has no obvious effect on LIF mRNA expression levels in RKO and HCT116 cells. (C) Ectopic HIF-2α expression increases LIF protein levels in RKO and HCT116 cells as determined by Western-blot assays. (D) and (E) HIF-2α binds to HRE-C and HRE-D sites in the LIF promoter in RKO cells transfected with HIF-2α expression plasmids as determined by ChIP assays. The amount of DNA fragments pulled-down was determined by real-time PCR (D) or conventional PCR (E) . The HRE site in the VEGF promoter serves as a positive control. (F) HIF-2α activates luciferase activity of reporter vectors containing HRE-C or HRE-D sites in the LIF promoter in RKO cells transfected with HIF-2α expression plasmids. Luciferase reporter vectors containing the HRE site in the VEGF promoter was included as a positive control. Data are presented as mean ± SD ( n = 3). *: p
    Figure Legend Snippet: HIF-2α transcriptionally regulates LIF expression (A) Ectopic HIF-2α expression increases LIF and VEGF mRNA expression levels in RKO and HCT116 cells. The mRNA expression levels of LIF and VEGF were determined by Taqman real-time PCR and normalized with actin. (B) Ectopic HIF-1α expression increases VEGF mRNA expression levels but has no obvious effect on LIF mRNA expression levels in RKO and HCT116 cells. (C) Ectopic HIF-2α expression increases LIF protein levels in RKO and HCT116 cells as determined by Western-blot assays. (D) and (E) HIF-2α binds to HRE-C and HRE-D sites in the LIF promoter in RKO cells transfected with HIF-2α expression plasmids as determined by ChIP assays. The amount of DNA fragments pulled-down was determined by real-time PCR (D) or conventional PCR (E) . The HRE site in the VEGF promoter serves as a positive control. (F) HIF-2α activates luciferase activity of reporter vectors containing HRE-C or HRE-D sites in the LIF promoter in RKO cells transfected with HIF-2α expression plasmids. Luciferase reporter vectors containing the HRE site in the VEGF promoter was included as a positive control. Data are presented as mean ± SD ( n = 3). *: p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Transfection, Chromatin Immunoprecipitation, Polymerase Chain Reaction, Positive Control, Luciferase, Activity Assay

    HIF-2α mediates the induction of LIF expression by hypoxia Knockdown of endogenous HIF-2α but not HIF-1α largely abolishes the induction of LIF expression by hypoxia in RKO cells. Cells with knockdown of endogenous HIF-1α, HIF-2α by siRNA oligos or transfected with control siRNA were treated with hypoxia for 36 h. Two different siRNA oligos against HIF-1α and HIF-2α, respectively, were used, and similar results were obtained. (A) The mRNA expression levels of LIF (left panel) and VEGF (right panel) were determined by Taqman real-time PCR and normalized with actin. (B) The LIF protein levels were determined by Western-blot assays. (C) The knockdown of HIF-1α (left panel) and HIF-2α (right panel) in cells was confirmed at the mRNA level by Taqman real-time PCR and normalized with actin. Data are presented as mean ± SD ( n = 3). *: p
    Figure Legend Snippet: HIF-2α mediates the induction of LIF expression by hypoxia Knockdown of endogenous HIF-2α but not HIF-1α largely abolishes the induction of LIF expression by hypoxia in RKO cells. Cells with knockdown of endogenous HIF-1α, HIF-2α by siRNA oligos or transfected with control siRNA were treated with hypoxia for 36 h. Two different siRNA oligos against HIF-1α and HIF-2α, respectively, were used, and similar results were obtained. (A) The mRNA expression levels of LIF (left panel) and VEGF (right panel) were determined by Taqman real-time PCR and normalized with actin. (B) The LIF protein levels were determined by Western-blot assays. (C) The knockdown of HIF-1α (left panel) and HIF-2α (right panel) in cells was confirmed at the mRNA level by Taqman real-time PCR and normalized with actin. Data are presented as mean ± SD ( n = 3). *: p

    Techniques Used: Expressing, Transfection, Real-time Polymerase Chain Reaction, Western Blot

    4) Product Images from "KPNA2 promotes metabolic reprogramming in glioblastomas by regulation of c-myc"

    Article Title: KPNA2 promotes metabolic reprogramming in glioblastomas by regulation of c-myc

    Journal: Journal of Experimental & Clinical Cancer Research : CR

    doi: 10.1186/s13046-018-0861-9

    KPNA2 promoted the glycolytic metabolism in the glioblastoma cells. a Levels of KPNA2 were analyzed by qRT-PCR and immunoblotting in the U87 glioblastoma cells transfected with the lentiviruses expressing small hairpin RNAs of KPNA2 (shKPNA2), wild-type KPNA2 (KPNA2), or a vector with scrambled nonspecific shRNAs(SC). NC is for the original U87 cells that without any handles, GAPDH served as a loading control. b mRNA levels and c enzymatic activities of HK2, PKM2 and PFK1 were determined in the U87 cells transfected with KPNA2-shRNA(siKPNA2), scrambled shRNA(SC) or wild-type KPNA2(KPNA2). Each bar represented the mean ± s.d. from three independent experiments. * P
    Figure Legend Snippet: KPNA2 promoted the glycolytic metabolism in the glioblastoma cells. a Levels of KPNA2 were analyzed by qRT-PCR and immunoblotting in the U87 glioblastoma cells transfected with the lentiviruses expressing small hairpin RNAs of KPNA2 (shKPNA2), wild-type KPNA2 (KPNA2), or a vector with scrambled nonspecific shRNAs(SC). NC is for the original U87 cells that without any handles, GAPDH served as a loading control. b mRNA levels and c enzymatic activities of HK2, PKM2 and PFK1 were determined in the U87 cells transfected with KPNA2-shRNA(siKPNA2), scrambled shRNA(SC) or wild-type KPNA2(KPNA2). Each bar represented the mean ± s.d. from three independent experiments. * P

    Techniques Used: Quantitative RT-PCR, Transfection, Expressing, Plasmid Preparation, shRNA

    5) Product Images from "MiR-196a exerts its oncogenic effect in glioblastoma multiforme by inhibition of IκBα both in vitro and in vivo"

    Article Title: MiR-196a exerts its oncogenic effect in glioblastoma multiforme by inhibition of IκBα both in vitro and in vivo

    Journal: Neuro-Oncology

    doi: 10.1093/neuonc/not307

    Effect of miR-196a on cell proliferation and apoptosis in vitro. (A) miR-196a expression was quantified by qRT–PCR in U87MG and T98G cells 48 h after transfection of miR-196a mimics, AMO-miR-196a, or negative control oligo. (B) Cell viability
    Figure Legend Snippet: Effect of miR-196a on cell proliferation and apoptosis in vitro. (A) miR-196a expression was quantified by qRT–PCR in U87MG and T98G cells 48 h after transfection of miR-196a mimics, AMO-miR-196a, or negative control oligo. (B) Cell viability

    Techniques Used: In Vitro, Expressing, Quantitative RT-PCR, Transfection, Negative Control

    6) Product Images from "Transcriptional Regulation of N-acetylaspartate Metabolism in the 5xFAD Model of Alzheimer's Disease: Evidence for Neuron-Glia Communication During Energetic Crisis"

    Article Title: Transcriptional Regulation of N-acetylaspartate Metabolism in the 5xFAD Model of Alzheimer's Disease: Evidence for Neuron-Glia Communication During Energetic Crisis

    Journal: Molecular and cellular neurosciences

    doi: 10.1016/j.mcn.2015.03.009

    ( A ) In-house QRT-PCR confirmation of the most significantly downregulated Complex I gene, Ndusf2 , confirms 4-month array data (5xFAD values expressed as fold-2 month wild type values, +/- SEM, n=5. ** p≤0.01). Reduced Ndusf2 expression correlates
    Figure Legend Snippet: ( A ) In-house QRT-PCR confirmation of the most significantly downregulated Complex I gene, Ndusf2 , confirms 4-month array data (5xFAD values expressed as fold-2 month wild type values, +/- SEM, n=5. ** p≤0.01). Reduced Ndusf2 expression correlates

    Techniques Used: Quantitative RT-PCR, Expressing

    ( A ) Increased aspa expression in 5xFAD brains at 2 months of age relative to wild type as assessed by QRT-PCR. Aspa expression presented as fold-wild type 2 month levels for each group, mean +/- SEM, n=5. Immunhistochemical evidence of increases in ASPA
    Figure Legend Snippet: ( A ) Increased aspa expression in 5xFAD brains at 2 months of age relative to wild type as assessed by QRT-PCR. Aspa expression presented as fold-wild type 2 month levels for each group, mean +/- SEM, n=5. Immunhistochemical evidence of increases in ASPA

    Techniques Used: Expressing, Quantitative RT-PCR

    Whole brain homogenates analyzed by HPLC show a significant reduction in NAA from 2-4 months of age in 5xFAD brains ( 1A ; mean +/- SEM shown for each genotype at each age, n=7). QRT-PCR analysis of Nat8L expression 2-4 months of age showing a significant
    Figure Legend Snippet: Whole brain homogenates analyzed by HPLC show a significant reduction in NAA from 2-4 months of age in 5xFAD brains ( 1A ; mean +/- SEM shown for each genotype at each age, n=7). QRT-PCR analysis of Nat8L expression 2-4 months of age showing a significant

    Techniques Used: High Performance Liquid Chromatography, Quantitative RT-PCR, Expressing

    7) Product Images from "Antimelanogenic effect of c-phycocyanin through modulation of tyrosinase expression by upregulation of ERK and downregulation of p38 MAPK signaling pathways"

    Article Title: Antimelanogenic effect of c-phycocyanin through modulation of tyrosinase expression by upregulation of ERK and downregulation of p38 MAPK signaling pathways

    Journal: Journal of Biomedical Science

    doi: 10.1186/1423-0127-18-74

    Cpc attenuated α-MSH-stimulated melanogenesis and elevated the abundance of intracellular cAMP . Cells were pretreated with 20 nM α-MSH for 30 mins, and then treated with Cpc (0.05, 0.1, 0.2 mg/mL) for 72 hrs. (A) Tyrosinase activity (black) and melanin content (grey) were measured. (B) The expression of tyrosinase was determined by immunoblotting analysis (black) and RT-PCR (grey), using β-actin and GAPDH as internal standards, respectively. (C) The cAMP concentration was measured by enzyme immunoassay at assigned time intervals (10, 30, 60 min) after Cpc treatment. Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P
    Figure Legend Snippet: Cpc attenuated α-MSH-stimulated melanogenesis and elevated the abundance of intracellular cAMP . Cells were pretreated with 20 nM α-MSH for 30 mins, and then treated with Cpc (0.05, 0.1, 0.2 mg/mL) for 72 hrs. (A) Tyrosinase activity (black) and melanin content (grey) were measured. (B) The expression of tyrosinase was determined by immunoblotting analysis (black) and RT-PCR (grey), using β-actin and GAPDH as internal standards, respectively. (C) The cAMP concentration was measured by enzyme immunoassay at assigned time intervals (10, 30, 60 min) after Cpc treatment. Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P

    Techniques Used: Activity Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Concentration Assay, Enzyme-linked Immunosorbent Assay

    Effect of Cpc on cAMP/MAPK/ERK pathway and MITF expression at protein and mRNA levels . Immunoblot analysis was performed with cell extract proteins treated with (A) Cpc (0.1 mg/mL) at assigned time intervals for ERK1/2 (control (black); CPC-treated (grey)), and (B) different Cpc concentration (0.05, 0.1, 0.2 mg/mL) at 540 min for MEK. (C) Cell extract proteins at assigned time intervals treated with Cpc (0.1 mg/mL) were examined by Immunoblot analysis for MITF using β-actin as internal standards (control (black); CPC-treated (grey)). (D) Different levels of Cpc (0.05, 0.1, 0.2 mg/mL) treated MITF mRNA were analyzed by Q-PCR at 540 min. (E) Immunoblot analysis treated with Cpc (0.1 mg/mL), PD98059 (PD, 20 μM), and Cpc+PD at 72 hrs were performed for the evaluation of MITF and tyrosinase expression (MITF (black); tyrosinase (grey)). Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P
    Figure Legend Snippet: Effect of Cpc on cAMP/MAPK/ERK pathway and MITF expression at protein and mRNA levels . Immunoblot analysis was performed with cell extract proteins treated with (A) Cpc (0.1 mg/mL) at assigned time intervals for ERK1/2 (control (black); CPC-treated (grey)), and (B) different Cpc concentration (0.05, 0.1, 0.2 mg/mL) at 540 min for MEK. (C) Cell extract proteins at assigned time intervals treated with Cpc (0.1 mg/mL) were examined by Immunoblot analysis for MITF using β-actin as internal standards (control (black); CPC-treated (grey)). (D) Different levels of Cpc (0.05, 0.1, 0.2 mg/mL) treated MITF mRNA were analyzed by Q-PCR at 540 min. (E) Immunoblot analysis treated with Cpc (0.1 mg/mL), PD98059 (PD, 20 μM), and Cpc+PD at 72 hrs were performed for the evaluation of MITF and tyrosinase expression (MITF (black); tyrosinase (grey)). Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P

    Techniques Used: Expressing, Concentration Assay, Polymerase Chain Reaction

    Effect of Cpc on viability of B16F10 melanoma cell, tyrosinase activity and melanin contents . Cells were treated with Cpc (0.05, 0.1, 0.2 mg/mL) for 72 hrs. (A) Cell viability was determined by MTT assay as described in Materials and Methods. (B) Tyrosinase activity (black) and melanin content (grey) were measured. (C) The expression of tyrosinase was determined by immunoblotting analysis (black) and RT-PCR (grey), using β-actin and GAPDH as internal standards, respectively. Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P
    Figure Legend Snippet: Effect of Cpc on viability of B16F10 melanoma cell, tyrosinase activity and melanin contents . Cells were treated with Cpc (0.05, 0.1, 0.2 mg/mL) for 72 hrs. (A) Cell viability was determined by MTT assay as described in Materials and Methods. (B) Tyrosinase activity (black) and melanin content (grey) were measured. (C) The expression of tyrosinase was determined by immunoblotting analysis (black) and RT-PCR (grey), using β-actin and GAPDH as internal standards, respectively. Data were expressed at mean ± SD from three different experiments. The asterisk (*) indicates a significant difference from control group (*, P

    Techniques Used: Activity Assay, MTT Assay, Expressing, Reverse Transcription Polymerase Chain Reaction

    8) Product Images from "Involvement of Interleukin-6 and Androgen Receptor Signaling in Pancreatic Cancer"

    Article Title: Involvement of Interleukin-6 and Androgen Receptor Signaling in Pancreatic Cancer

    Journal: Genes & Cancer

    doi: 10.1177/1947601910383417

    Migration of pancreatic cancer cell lines was investigated using a wound-healing scratch assay ( A-E ). Knockdown of AR suppressed migration of pancreatic cancer cells in the presence of IL-6 and DHT ( F-H ). KP-2 ( A ), SUIT-2 ( B ), Panc-1 ( C ), AsPC-1 ( D ), and MIAPaCa-2 ( E ) cells were subjected to wound-healing scratch assay. Cells were incubated with or without IL-6 (100 ng/mL) and/or DHT (10 nM) for approximately 24 hours. The migration thus observed was presented as a ratio of migration, considering migration in untreated control as 1. Scion images were used for analysis. The error bars represent mean ± SD of 3 independent experiments. Results of semiquantitative RT-PCR analysis for AR and GAPDH mRNAs are shown in the lower part below each graph ( A-E ). KP-2 cells were transfected with 10 nM AR siRNA (si-AR) or 10 nM control siRNA (si-C) (Thermo Fisher Scientific Dharmacon), subjected to wound-healing scratch assay, and incubated with IL-6 (100 ng/mL) and DHT (10 nM) for 24 hours. ( F ) Light microscopy pictures show KP-2 cells transfected with 10 nM si-C or 10 nM si-AR and incubated with 100 ng/mL IL-6 and 10 nM DHT at 0 hours (left panel) and 24 hours later (right panels). ( G ) The migration observed was presented as a ratio of migration, considering migration in transfection with si-C as 1. The error bars represent mean ± SD of 3 independent experiments. ( H ) Knockdown of AR expression in KP-2 cells. Whole cell lysates from KP-2 cells after si-C or si-AR treatment were immunoblotted with antibodies against AR or GAPDH.
    Figure Legend Snippet: Migration of pancreatic cancer cell lines was investigated using a wound-healing scratch assay ( A-E ). Knockdown of AR suppressed migration of pancreatic cancer cells in the presence of IL-6 and DHT ( F-H ). KP-2 ( A ), SUIT-2 ( B ), Panc-1 ( C ), AsPC-1 ( D ), and MIAPaCa-2 ( E ) cells were subjected to wound-healing scratch assay. Cells were incubated with or without IL-6 (100 ng/mL) and/or DHT (10 nM) for approximately 24 hours. The migration thus observed was presented as a ratio of migration, considering migration in untreated control as 1. Scion images were used for analysis. The error bars represent mean ± SD of 3 independent experiments. Results of semiquantitative RT-PCR analysis for AR and GAPDH mRNAs are shown in the lower part below each graph ( A-E ). KP-2 cells were transfected with 10 nM AR siRNA (si-AR) or 10 nM control siRNA (si-C) (Thermo Fisher Scientific Dharmacon), subjected to wound-healing scratch assay, and incubated with IL-6 (100 ng/mL) and DHT (10 nM) for 24 hours. ( F ) Light microscopy pictures show KP-2 cells transfected with 10 nM si-C or 10 nM si-AR and incubated with 100 ng/mL IL-6 and 10 nM DHT at 0 hours (left panel) and 24 hours later (right panels). ( G ) The migration observed was presented as a ratio of migration, considering migration in transfection with si-C as 1. The error bars represent mean ± SD of 3 independent experiments. ( H ) Knockdown of AR expression in KP-2 cells. Whole cell lysates from KP-2 cells after si-C or si-AR treatment were immunoblotted with antibodies against AR or GAPDH.

    Techniques Used: Migration, Wound Healing Assay, Incubation, Reverse Transcription Polymerase Chain Reaction, Transfection, Light Microscopy, Expressing

    AR and IL-6R expression in pancreatic cancer cell lines. ( A ) Relative expression of AR mRNA. RNAs from MIAPaCa-2, Panc-1, AsPC-1, SUIT-2, KP-2 cells, and LNCaP (AR-positive) cells were subjected to real-time RT-PCR for AR and GAPDH gene expression. AR/GAPDH mRNA in MIAPaCa-2 was set as 1-fold. The results were based on the relative expression level of the housekeeping gene GAPDH as an internal control. ( B ) IL-6Rα, GP130, AR, and GAPDH protein expression. Total cell lysates (5 µg) of MIAPaCa-2, Panc-1, AsPC-1, SUIT-2, KP-2, and LNCaP were subjected to 10% SDS-PAGE for immunoblotting with specific antibodies against IL-6Rα (Santa Cruz Biotechnology), GP130 (Cell Signaling Technology), AR (Santa Cruz Biotechnology), and GAPDH (Santa Cruz Biotechnology).
    Figure Legend Snippet: AR and IL-6R expression in pancreatic cancer cell lines. ( A ) Relative expression of AR mRNA. RNAs from MIAPaCa-2, Panc-1, AsPC-1, SUIT-2, KP-2 cells, and LNCaP (AR-positive) cells were subjected to real-time RT-PCR for AR and GAPDH gene expression. AR/GAPDH mRNA in MIAPaCa-2 was set as 1-fold. The results were based on the relative expression level of the housekeeping gene GAPDH as an internal control. ( B ) IL-6Rα, GP130, AR, and GAPDH protein expression. Total cell lysates (5 µg) of MIAPaCa-2, Panc-1, AsPC-1, SUIT-2, KP-2, and LNCaP were subjected to 10% SDS-PAGE for immunoblotting with specific antibodies against IL-6Rα (Santa Cruz Biotechnology), GP130 (Cell Signaling Technology), AR (Santa Cruz Biotechnology), and GAPDH (Santa Cruz Biotechnology).

    Techniques Used: Expressing, Quantitative RT-PCR, SDS Page

    9) Product Images from "Clinical profile of hearing loss in children with congenital cytomegalovirus (CMV) infection: CMV DNA diagnosis using preserved umbilical cord"

    Article Title: Clinical profile of hearing loss in children with congenital cytomegalovirus (CMV) infection: CMV DNA diagnosis using preserved umbilical cord

    Journal: Acta Oto-Laryngologica

    doi: 10.3109/00016489.2011.583268

    An original amplification plot of real-time PCR in case no. 12 with positive CMV DNA. CMV DNA in positive control and case 12 (A) and genomic DNA (GJB2 gene) in positive control, case no. 12 and negative control (B) are amplified. These results show that our real-time PCR method is precise. Blank, sample without any added DNA.
    Figure Legend Snippet: An original amplification plot of real-time PCR in case no. 12 with positive CMV DNA. CMV DNA in positive control and case 12 (A) and genomic DNA (GJB2 gene) in positive control, case no. 12 and negative control (B) are amplified. These results show that our real-time PCR method is precise. Blank, sample without any added DNA.

    Techniques Used: Amplification, Real-time Polymerase Chain Reaction, Positive Control, Negative Control

    10) Product Images from "Divergent Roles for Airway Epithelial MMP7 and Retinoic Acid in Experimental Asthma"

    Article Title: Divergent Roles for Airway Epithelial MMP7 and Retinoic Acid in Experimental Asthma

    Journal: Nature immunology

    doi: 10.1038/ni.1719

    Attenuated T H 2 cytokines and chemokines in Mmp7 −/− mice Mmp7 −/− mice and age and sex matched wild-type (n=5 per group) mice immunized as described in Fig. 2 and 24 h after the last immunization, concentrations of (a) IL-4, (b) IL-5 and (c) IL-13 were measured in BAL fluid using Luminex. (d) Level of CCL11 in BAL fluid was measured by ELISA. (e) Expression of Il-25 mRNA in the lung of WT and Mmp7 −/− CAA immunized mice was measured by real time RT-PCR. Data was normalized to actin. (Data represent mean values ± SD; representative of 3 independent experiments). * P
    Figure Legend Snippet: Attenuated T H 2 cytokines and chemokines in Mmp7 −/− mice Mmp7 −/− mice and age and sex matched wild-type (n=5 per group) mice immunized as described in Fig. 2 and 24 h after the last immunization, concentrations of (a) IL-4, (b) IL-5 and (c) IL-13 were measured in BAL fluid using Luminex. (d) Level of CCL11 in BAL fluid was measured by ELISA. (e) Expression of Il-25 mRNA in the lung of WT and Mmp7 −/− CAA immunized mice was measured by real time RT-PCR. Data was normalized to actin. (Data represent mean values ± SD; representative of 3 independent experiments). * P

    Techniques Used: Mouse Assay, Luminex, Enzyme-linked Immunosorbent Assay, Expressing, Cellular Antioxidant Activity Assay, Quantitative RT-PCR

    11) Product Images from "Multiple Sclerosis Treatments Affect Monocyte-Derived Microvesicle Production"

    Article Title: Multiple Sclerosis Treatments Affect Monocyte-Derived Microvesicle Production

    Journal: Frontiers in Neurology

    doi: 10.3389/fneur.2017.00422

    Expression level of M1 markers evaluated by RT-PCR: interleukin-1β (IL-1B) (A) , TNFalpha (TNFa) (B) , IL-6 (C) , and IL-18 (D) in unstimulated Krebs-Ringer solution (KRH) or benzoyl-ATP (BzATP)-stimulated monocytes in MS patients at baseline and after 2, 6, and 12 months of treatments with interferon-beta (IFNb) and teriflunomide. Data are expressed as mean ± SEM of fold change values (* p
    Figure Legend Snippet: Expression level of M1 markers evaluated by RT-PCR: interleukin-1β (IL-1B) (A) , TNFalpha (TNFa) (B) , IL-6 (C) , and IL-18 (D) in unstimulated Krebs-Ringer solution (KRH) or benzoyl-ATP (BzATP)-stimulated monocytes in MS patients at baseline and after 2, 6, and 12 months of treatments with interferon-beta (IFNb) and teriflunomide. Data are expressed as mean ± SEM of fold change values (* p

    Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Mass Spectrometry

    12) Product Images from "Engineering Xeno-Free Microcarriers with Recombinant Vitronectin, Albumin and UV Irradiation for Human Pluripotent Stem Cell Bioprocessing"

    Article Title: Engineering Xeno-Free Microcarriers with Recombinant Vitronectin, Albumin and UV Irradiation for Human Pluripotent Stem Cell Bioprocessing

    Journal: ACS biomaterials science & engineering

    doi: 10.1021/acsbiomaterials.6b00253

    Embryoid body (EB) formation and multilineage differentiation for hPSCs expanded in VN+HSA+UV microcarrier cultures for 5 continuous passages. (A) Cells within EBs displayed markers of DE, MS and NE analyzed by qPCR. Expression was normalized to that of undifferentiated cells with ACTB as the housekeeping gene. Quantitative PCR analysis of hPSCs expanded for 5 passages in microcarrier cultures and subjected to directed differentiation toward (B) DE, (C) MS and (D) NE and the corresponding immunocytochemistry results (E–G). The expression results in (B–D) was normalized to that of hPSCs cultured in basal media without the differentiation factors.
    Figure Legend Snippet: Embryoid body (EB) formation and multilineage differentiation for hPSCs expanded in VN+HSA+UV microcarrier cultures for 5 continuous passages. (A) Cells within EBs displayed markers of DE, MS and NE analyzed by qPCR. Expression was normalized to that of undifferentiated cells with ACTB as the housekeeping gene. Quantitative PCR analysis of hPSCs expanded for 5 passages in microcarrier cultures and subjected to directed differentiation toward (B) DE, (C) MS and (D) NE and the corresponding immunocytochemistry results (E–G). The expression results in (B–D) was normalized to that of hPSCs cultured in basal media without the differentiation factors.

    Techniques Used: Mass Spectrometry, Real-time Polymerase Chain Reaction, Expressing, Immunocytochemistry, Cell Culture

    13) Product Images from "Identification of microRNA-regulated pathways using an integration of microRNA-mRNA microarray and bioinformatics analysis in CD34+ cells of myelodysplastic syndromes"

    Article Title: Identification of microRNA-regulated pathways using an integration of microRNA-mRNA microarray and bioinformatics analysis in CD34+ cells of myelodysplastic syndromes

    Journal: Scientific Reports

    doi: 10.1038/srep32232

    Validation of screened miRNAs and mRNAs using qRT-PCR. The expression of miR-17-3p ( A ) and miR-19a-3p ( B ) were significantly higher in the high-grade and low-grade MDS groups than those in the normal control group (all P
    Figure Legend Snippet: Validation of screened miRNAs and mRNAs using qRT-PCR. The expression of miR-17-3p ( A ) and miR-19a-3p ( B ) were significantly higher in the high-grade and low-grade MDS groups than those in the normal control group (all P

    Techniques Used: Quantitative RT-PCR, Expressing

    14) Product Images from "Exploring the Mechanisms of Electroacupuncture-Induced Analgesia through RNA Sequencing of the Periaqueductal Gray"

    Article Title: Exploring the Mechanisms of Electroacupuncture-Induced Analgesia through RNA Sequencing of the Periaqueductal Gray

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms19010002

    qRT-PCR validation of DEGs. The reliability of the sequencing data was validated by qRT-PCR. The change tendency of fifteen DEGs was consistent with the results from sequencing data. The data depicted by the y -axis were calculated using the expression values of 2 −ΔΔ C t and expressed as the mean ± SD in triplicate. The significance of differences in expression between samples was calculated by a t test. * Compared with the control p
    Figure Legend Snippet: qRT-PCR validation of DEGs. The reliability of the sequencing data was validated by qRT-PCR. The change tendency of fifteen DEGs was consistent with the results from sequencing data. The data depicted by the y -axis were calculated using the expression values of 2 −ΔΔ C t and expressed as the mean ± SD in triplicate. The significance of differences in expression between samples was calculated by a t test. * Compared with the control p

    Techniques Used: Quantitative RT-PCR, Sequencing, Expressing

    15) Product Images from "Ribose Accelerates Gut Motility and Suppresses Mouse Body Weight Gaining"

    Article Title: Ribose Accelerates Gut Motility and Suppresses Mouse Body Weight Gaining

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.13635

    Expression pattern of Rbks gene in mice. (A) Quantitative real-time (qRT) PCR revealed a high RNA level of Rbks gene in kidney, liver, testis, small intestine (duodenum, jejunum and ileum) and cecum of wild type mice. Primers for qRT PCR located on exon5 and exon6. (B) The expression pattern of Rbks gene was also revealed by whole mount X-gal staining with Rbks galeo/galeo mice tissues. The wild type litter mates served as controls. Black bar: 1cm. (C) The whole mount staining intestines were further processed for paraffin sections, and the cellular β-galactosidase signals were identified in intestinal brush boarders. Black bar: 100μm.
    Figure Legend Snippet: Expression pattern of Rbks gene in mice. (A) Quantitative real-time (qRT) PCR revealed a high RNA level of Rbks gene in kidney, liver, testis, small intestine (duodenum, jejunum and ileum) and cecum of wild type mice. Primers for qRT PCR located on exon5 and exon6. (B) The expression pattern of Rbks gene was also revealed by whole mount X-gal staining with Rbks galeo/galeo mice tissues. The wild type litter mates served as controls. Black bar: 1cm. (C) The whole mount staining intestines were further processed for paraffin sections, and the cellular β-galactosidase signals were identified in intestinal brush boarders. Black bar: 100μm.

    Techniques Used: Expressing, Mouse Assay, Quantitative RT-PCR, Staining

    16) Product Images from "Genetic profile and biological implication of PIN2/TRF1-interacting telomerase inhibitor 1 (PinX1) in human cancers: an analysis using The Cancer Genome Atlas"

    Article Title: Genetic profile and biological implication of PIN2/TRF1-interacting telomerase inhibitor 1 (PinX1) in human cancers: an analysis using The Cancer Genome Atlas

    Journal: Oncotarget

    doi: 10.18632/oncotarget.18589

    qRT-PCR used to analyze PinX1 mRNA expression status in nine types of human cancers Nine types of human cancer tissues were selected to analyze PinX1 mRNA expression using qRT-PCR. Low PinX1 mRNA expression was observed in 9 out of 12 (75%) prostate adenocarcinoma patient samples (A) , 8 out of 12 (66.7%) colorectal adenocarcinoma samples (B) , 7 out of 12 (58.3%) head and neck squamous cell carcinoma samples (C) , 7 out of 12 (58.3%) kidney renal clear cell carcinoma samples (D) , 11 out of 12 (91.7%) lung adenocarcinoma samples (E) , 8 out of 12 (66.7%) lung squamous cell carcinoma samples (F) , 8 out of 12 (66.7%) bladder urothelial carcinoma samples (G) , 8 out of 12 (66.7%) ovarian serous cystadenocarcinoma samples (H) , 6 out of 12 (50%) esophagus carcinoma tissues samples (I) . All cancerous tissue was compared to normal tissue samples.
    Figure Legend Snippet: qRT-PCR used to analyze PinX1 mRNA expression status in nine types of human cancers Nine types of human cancer tissues were selected to analyze PinX1 mRNA expression using qRT-PCR. Low PinX1 mRNA expression was observed in 9 out of 12 (75%) prostate adenocarcinoma patient samples (A) , 8 out of 12 (66.7%) colorectal adenocarcinoma samples (B) , 7 out of 12 (58.3%) head and neck squamous cell carcinoma samples (C) , 7 out of 12 (58.3%) kidney renal clear cell carcinoma samples (D) , 11 out of 12 (91.7%) lung adenocarcinoma samples (E) , 8 out of 12 (66.7%) lung squamous cell carcinoma samples (F) , 8 out of 12 (66.7%) bladder urothelial carcinoma samples (G) , 8 out of 12 (66.7%) ovarian serous cystadenocarcinoma samples (H) , 6 out of 12 (50%) esophagus carcinoma tissues samples (I) . All cancerous tissue was compared to normal tissue samples.

    Techniques Used: Quantitative RT-PCR, Expressing

    17) Product Images from "Prolonged oxidative stress down-regulates Early B cell factor 1 with inhibition of its tumor suppressive function against cholangiocarcinoma genesis"

    Article Title: Prolonged oxidative stress down-regulates Early B cell factor 1 with inhibition of its tumor suppressive function against cholangiocarcinoma genesis

    Journal: Redox Biology

    doi: 10.1016/j.redox.2017.11.011

    (A) Relative mRNA expression levels of TFF1 was measured by real-time PCR and adjusted by β-actin mRNA expression. (B) The graphical data represent the migrated cells detected by the migration assay. The Y axis represents the number of migrating cells per field and the X axis the experimental group. The asterisk (*) indicates statistical significance at P
    Figure Legend Snippet: (A) Relative mRNA expression levels of TFF1 was measured by real-time PCR and adjusted by β-actin mRNA expression. (B) The graphical data represent the migrated cells detected by the migration assay. The Y axis represents the number of migrating cells per field and the X axis the experimental group. The asterisk (*) indicates statistical significance at P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Migration

    (A) Relative mRNA expression levels of EBF1 was measured by real-time PCR and adjusted by β-actin mRNA expression in MMNK1, ox-MMNK1-L and CCA cell lines. The symbol asterisk (*) indicates statistical significance at P
    Figure Legend Snippet: (A) Relative mRNA expression levels of EBF1 was measured by real-time PCR and adjusted by β-actin mRNA expression in MMNK1, ox-MMNK1-L and CCA cell lines. The symbol asterisk (*) indicates statistical significance at P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    Relative mRNA expression levels of EBF1 (A), CD133 (B) and Oct3/4 (C) were measured by real-time PCR and adjusted by β-actin mRNA expression. The asterisk (*) indicates statistical significance at P
    Figure Legend Snippet: Relative mRNA expression levels of EBF1 (A), CD133 (B) and Oct3/4 (C) were measured by real-time PCR and adjusted by β-actin mRNA expression. The asterisk (*) indicates statistical significance at P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    18) Product Images from "N-Myc-induced up-regulation of TRPM6/TRPM7 channels promotes neuroblastoma cell proliferation"

    Article Title: N-Myc-induced up-regulation of TRPM6/TRPM7 channels promotes neuroblastoma cell proliferation

    Journal: Oncotarget

    doi:

    TRPM6 and TRPM7 correlation with MYCN in neuroblastoma A-C, N-Myc, TRPM7 and TRPM6 mRNA expression correlation with MYCN amplification in the Kocak-649 cohort. Microarray analysis of N-Myc, TRPM7 and TRPM6 mRNA expression in Kocak-649 (GSE45547), the largest neuroblastoma cohort in the public domain. The graphs present N-Myc (A), TRPM7 (B) and TRPM6 (C) expression in tumors without (n=550) and with (n=93) MYCN amplification. Y-axes represent sample ranks in a non-parametric Kruskal-Wallis t test; actual mean ± s.e.m. expression values were: 17,839 ± 1,137 (MYCN), 104.9 ± 3.7 (TRPM6), 807.0 ± 321.1 (TRPM7). Both N-Myc and TRPM7 expression are significantly higher in tumors with MYCN amplification ( p =1.5 • 10 −51 and p =2.3 • 10 −7 respectively; Kruskal-Wallis t test). D-F, qRT-PCR analysis of N-Myc (D), TRPM7 (E) and TRPM6 (F) expression levels in the SHEP-21N cell line where MYCN transgene is controlled by tetracycline, i.e., N-Myc expression is repressed in the presence of tetracycline (control), but removal of tetracycline induces N-Myc expression (N-Myc). The graphs show normalized N-Myc, TRPM7 and TRPM6 expression in SHEP-21N cells with (N-Myc, n = 9) or without (control, n = 9) N-Myc expression. *, p
    Figure Legend Snippet: TRPM6 and TRPM7 correlation with MYCN in neuroblastoma A-C, N-Myc, TRPM7 and TRPM6 mRNA expression correlation with MYCN amplification in the Kocak-649 cohort. Microarray analysis of N-Myc, TRPM7 and TRPM6 mRNA expression in Kocak-649 (GSE45547), the largest neuroblastoma cohort in the public domain. The graphs present N-Myc (A), TRPM7 (B) and TRPM6 (C) expression in tumors without (n=550) and with (n=93) MYCN amplification. Y-axes represent sample ranks in a non-parametric Kruskal-Wallis t test; actual mean ± s.e.m. expression values were: 17,839 ± 1,137 (MYCN), 104.9 ± 3.7 (TRPM6), 807.0 ± 321.1 (TRPM7). Both N-Myc and TRPM7 expression are significantly higher in tumors with MYCN amplification ( p =1.5 • 10 −51 and p =2.3 • 10 −7 respectively; Kruskal-Wallis t test). D-F, qRT-PCR analysis of N-Myc (D), TRPM7 (E) and TRPM6 (F) expression levels in the SHEP-21N cell line where MYCN transgene is controlled by tetracycline, i.e., N-Myc expression is repressed in the presence of tetracycline (control), but removal of tetracycline induces N-Myc expression (N-Myc). The graphs show normalized N-Myc, TRPM7 and TRPM6 expression in SHEP-21N cells with (N-Myc, n = 9) or without (control, n = 9) N-Myc expression. *, p

    Techniques Used: Expressing, Amplification, Microarray, Quantitative RT-PCR

    Endogenous MagNuM currents are mediated by TRPM7 and TRPM6 Currents were measured in divalent-free internal solution with 5 mM EGTA and 5 mM EDTA. A-B, siRNAs efficacy measured by qRT-PCR analysis of TRPM6 (A) and TRPM7 (B) expression levels in the SHEP-21N cell line under both control and N-Myc upregulation conditions (n = 9). C, MagNuM currents in control SHEP-21N cells treated with negative non-silencing (siCTL) or specific siRNA sequences against TRPM7, TRPM6, TRPM7 TRPM6. D, current measurement in N-Myc-expressing SHEP-21N cells treated with negative (siCTL) or specific siRNA sequences against TRPM7, TRPM6, TRPM7 TRPM6. E, statistical summary of current amplitudes at 700 s as in c-d (*, p
    Figure Legend Snippet: Endogenous MagNuM currents are mediated by TRPM7 and TRPM6 Currents were measured in divalent-free internal solution with 5 mM EGTA and 5 mM EDTA. A-B, siRNAs efficacy measured by qRT-PCR analysis of TRPM6 (A) and TRPM7 (B) expression levels in the SHEP-21N cell line under both control and N-Myc upregulation conditions (n = 9). C, MagNuM currents in control SHEP-21N cells treated with negative non-silencing (siCTL) or specific siRNA sequences against TRPM7, TRPM6, TRPM7 TRPM6. D, current measurement in N-Myc-expressing SHEP-21N cells treated with negative (siCTL) or specific siRNA sequences against TRPM7, TRPM6, TRPM7 TRPM6. E, statistical summary of current amplitudes at 700 s as in c-d (*, p

    Techniques Used: Quantitative RT-PCR, Expressing

    19) Product Images from "RhMYB108, an R2R3-MYB transcription factor, is involved in ethylene- and JA-induced petal senescence in rose plants"

    Article Title: RhMYB108, an R2R3-MYB transcription factor, is involved in ethylene- and JA-induced petal senescence in rose plants

    Journal: Horticulture Research

    doi: 10.1038/s41438-019-0221-8

    RhMYB108 directly binds to the promoter of RhNAC053, RhNAC092 and RhSAG113 . a qRT-PCR analysis of SAG expression in RhMYB108- silenced petals. b Interaction of RhMYB108 protein with the SAG promoter regions, as revealed using yeast one-hybrid assays. Interactions were determined based on yeast cell growth and were confirmed by the color indication of X-β-gal on SD/-Trp/-Ura medium plates. c , d Images of firefly luciferase fluorescence signals and relative reporter activity (LUC/REN) in N. benthamiana leaves. RhMYB108 protein was separately coexpressed with pNAC053::LUC , pNAC092::LUC and pSAG113::LUC in tobacco leaves. After 48 h of Agrobacterium tumefaciens infiltration, live LUC images and relative LUC activity (LUC/REN) in tobacco leaves were assayed. The error bars represent the means of six biological replicates, and the asterisks indicate statistically significant differences according to Student’s t test (* P
    Figure Legend Snippet: RhMYB108 directly binds to the promoter of RhNAC053, RhNAC092 and RhSAG113 . a qRT-PCR analysis of SAG expression in RhMYB108- silenced petals. b Interaction of RhMYB108 protein with the SAG promoter regions, as revealed using yeast one-hybrid assays. Interactions were determined based on yeast cell growth and were confirmed by the color indication of X-β-gal on SD/-Trp/-Ura medium plates. c , d Images of firefly luciferase fluorescence signals and relative reporter activity (LUC/REN) in N. benthamiana leaves. RhMYB108 protein was separately coexpressed with pNAC053::LUC , pNAC092::LUC and pSAG113::LUC in tobacco leaves. After 48 h of Agrobacterium tumefaciens infiltration, live LUC images and relative LUC activity (LUC/REN) in tobacco leaves were assayed. The error bars represent the means of six biological replicates, and the asterisks indicate statistically significant differences according to Student’s t test (* P

    Techniques Used: Quantitative RT-PCR, Expressing, Luciferase, Fluorescence, Activity Assay

    qRT-PCR analysis of the expression level of RhMYB108 . a Expression profiles of RhMYB108 in the roots, stems, leaves, and petals of rose plants. b , c , d , e , f Expression profiles of RhMYB108 in the petals, receptacles, sepals, stamens, and pistils at various stages of flower opening. g Expression of RhMYB108 in response to exogenous hormones. ETH, 10 μL/L ethylene; JA, 100 μM MeJA; 1-MCP, 2 μL/L. The petals at stage 2 were used as materials in g . RhUBI2 was used as an internal control. Three independent biological repeats were tested, and the data are presented as the means ± SDs. The letters indicate significant differences according to Duncan’s multiple range test ( P
    Figure Legend Snippet: qRT-PCR analysis of the expression level of RhMYB108 . a Expression profiles of RhMYB108 in the roots, stems, leaves, and petals of rose plants. b , c , d , e , f Expression profiles of RhMYB108 in the petals, receptacles, sepals, stamens, and pistils at various stages of flower opening. g Expression of RhMYB108 in response to exogenous hormones. ETH, 10 μL/L ethylene; JA, 100 μM MeJA; 1-MCP, 2 μL/L. The petals at stage 2 were used as materials in g . RhUBI2 was used as an internal control. Three independent biological repeats were tested, and the data are presented as the means ± SDs. The letters indicate significant differences according to Duncan’s multiple range test ( P

    Techniques Used: Quantitative RT-PCR, Expressing

    20) Product Images from "Atrasentan increased the expression of klotho by mediating miR-199b-5p and prevented renal tubular injury in diabetic nephropathy"

    Article Title: Atrasentan increased the expression of klotho by mediating miR-199b-5p and prevented renal tubular injury in diabetic nephropathy

    Journal: Scientific Reports

    doi: 10.1038/srep19979

    Atrasentan up-regulated miR-199b-5p expression, down-regulated klotho expression, and increased the antioxidant ability of HK-2 cells exposed to a high concentration of glucose. The HK-2 cells were incubated with different concentrations of glucose (5 mmol/L or 20 mmol/L) for 72 h and treated with atrasentan. The expression of miR-199b-5p was quantified by real-time PCR ( A ). The expression of klotho was measured by real-time PCR and a Western blot ( B ). Levels of antioxidant indicators (T-AOC, SOD, CAT, and GSH) of renal mitochondria in HK-2 cells were detected by commercial kits ( C–F ). * P
    Figure Legend Snippet: Atrasentan up-regulated miR-199b-5p expression, down-regulated klotho expression, and increased the antioxidant ability of HK-2 cells exposed to a high concentration of glucose. The HK-2 cells were incubated with different concentrations of glucose (5 mmol/L or 20 mmol/L) for 72 h and treated with atrasentan. The expression of miR-199b-5p was quantified by real-time PCR ( A ). The expression of klotho was measured by real-time PCR and a Western blot ( B ). Levels of antioxidant indicators (T-AOC, SOD, CAT, and GSH) of renal mitochondria in HK-2 cells were detected by commercial kits ( C–F ). * P

    Techniques Used: Expressing, Concentration Assay, Incubation, Real-time Polymerase Chain Reaction, Western Blot

    miR-199b-5p targeted klotho and regulated its expression in HK-2 cells. The HK-2 cells were transfected with an miR-199b-5p mimic/inhibitor or NC, and the expression of miR-199b-5p was quantified by real-time PCR ( A,E ). The 3′ UTR activity of klotho was detected by a luciferase reporter assay ( B,F ). The expression of klotho was quantified by real-time PCR and a Western blot ( C,G ). The klotho concentration of cell culture supernatant was determined with an ELISA kit ( D , H ). * P
    Figure Legend Snippet: miR-199b-5p targeted klotho and regulated its expression in HK-2 cells. The HK-2 cells were transfected with an miR-199b-5p mimic/inhibitor or NC, and the expression of miR-199b-5p was quantified by real-time PCR ( A,E ). The 3′ UTR activity of klotho was detected by a luciferase reporter assay ( B,F ). The expression of klotho was quantified by real-time PCR and a Western blot ( C,G ). The klotho concentration of cell culture supernatant was determined with an ELISA kit ( D , H ). * P

    Techniques Used: Expressing, Transfection, Real-time Polymerase Chain Reaction, Activity Assay, Luciferase, Reporter Assay, Western Blot, Concentration Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

    Atrasentan decreased histone deacetylation of the miR-199b-5p promoter in HK-2 cells exposed to high glucose. The HK-2 cells were exposed to various concentrations of TSA (0.1, 0.5 and 1.0 μmol/L) for 72 h and the expression of miR-199b-5p was then quantified by real-time PCR ( A ). The HK-2 cells were incubated with various concentrations of glucose (5 mmol/L or 20 mmol/L) and treated with atrasentan. DNA-H3 and DNA-H4 complex levels were detected by a ChIP assay ( C , D ). * P
    Figure Legend Snippet: Atrasentan decreased histone deacetylation of the miR-199b-5p promoter in HK-2 cells exposed to high glucose. The HK-2 cells were exposed to various concentrations of TSA (0.1, 0.5 and 1.0 μmol/L) for 72 h and the expression of miR-199b-5p was then quantified by real-time PCR ( A ). The HK-2 cells were incubated with various concentrations of glucose (5 mmol/L or 20 mmol/L) and treated with atrasentan. DNA-H3 and DNA-H4 complex levels were detected by a ChIP assay ( C , D ). * P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Incubation, Chromatin Immunoprecipitation

    miR-199b-5p participated in the effects of atrasentan on the expression of klotho, antioxidant activities, and caspase activity of HK-2 cells. The HK-2 cells were treated with 20 mmol/L glucose and co-incubated with atrasentan. An miR-199b-5p mimic (100 mmol) or an NC was then transfected into the cells. The concentration of klotho treated in cells was 20 nmol/L. The expression of klotho was measured by real-time PCR and a Western blot ( A ). Levels of antioxidant indicators (T-AOC, SOD, CAT, and GSH) of renal mitochondria were detected by commercial kits in the HK-2 cells ( B–E ). The activity of caspase-3 was detected with an ELISA kit ( F ). * P
    Figure Legend Snippet: miR-199b-5p participated in the effects of atrasentan on the expression of klotho, antioxidant activities, and caspase activity of HK-2 cells. The HK-2 cells were treated with 20 mmol/L glucose and co-incubated with atrasentan. An miR-199b-5p mimic (100 mmol) or an NC was then transfected into the cells. The concentration of klotho treated in cells was 20 nmol/L. The expression of klotho was measured by real-time PCR and a Western blot ( A ). Levels of antioxidant indicators (T-AOC, SOD, CAT, and GSH) of renal mitochondria were detected by commercial kits in the HK-2 cells ( B–E ). The activity of caspase-3 was detected with an ELISA kit ( F ). * P

    Techniques Used: Expressing, Activity Assay, Incubation, Transfection, Concentration Assay, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay

    Atrasentan improved renal function, decreased miR-199b-5p expression, and increased klotho expression in STZ-induced DN mice. The urinary albumin/creatinine ( A ), serum BUN (blood urea nitrogen) ( B ), and serum creatinine ( C ) were measured to evaluate renal function. Real-time PCR was used to detect serum and renal tubular epithelial cell miR-199-5p expression ( D ). The serum klotho level was measured with an ELISA kit, and the renal klotho level was detected with real-time PCR and a Western blot ( E ). The levels of urinary KIM-1(kidney injury molecule-1) ( F ), NGAL (neutrophil gelatinase-associated lipocalin) ( G ) and NAG (N-acetyl-β-D glucosminidase) ( H ) were measured by ELISA kits. A Caspase-3 activity assay ( F ) and a TUNEL assay ( G ) were used to assess the apoptosis of renal tubular epithelial cells of mice. n = 10. * P
    Figure Legend Snippet: Atrasentan improved renal function, decreased miR-199b-5p expression, and increased klotho expression in STZ-induced DN mice. The urinary albumin/creatinine ( A ), serum BUN (blood urea nitrogen) ( B ), and serum creatinine ( C ) were measured to evaluate renal function. Real-time PCR was used to detect serum and renal tubular epithelial cell miR-199-5p expression ( D ). The serum klotho level was measured with an ELISA kit, and the renal klotho level was detected with real-time PCR and a Western blot ( E ). The levels of urinary KIM-1(kidney injury molecule-1) ( F ), NGAL (neutrophil gelatinase-associated lipocalin) ( G ) and NAG (N-acetyl-β-D glucosminidase) ( H ) were measured by ELISA kits. A Caspase-3 activity assay ( F ) and a TUNEL assay ( G ) were used to assess the apoptosis of renal tubular epithelial cells of mice. n = 10. * P

    Techniques Used: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot, Caspase-3 Activity Assay, TUNEL Assay

    21) Product Images from "Broomrape infestation in carrot (Daucus carota): Changes in carotenoid gene expression and carotenoid accumulation in the parasitic weed Phelipanche aegyptiaca and its host"

    Article Title: Broomrape infestation in carrot (Daucus carota): Changes in carotenoid gene expression and carotenoid accumulation in the parasitic weed Phelipanche aegyptiaca and its host

    Journal: Scientific Reports

    doi: 10.1038/s41598-019-57298-7

    qRT-PCR determination of transcript levels of carrots ( D . carota ) DcPSY1 , DcCRTISO , DcD27 , DcCCD7 , and DcCCD8 in roots of different carrot cultivars before and after the infestion by obligate parasite P . aegyptiaca . Quantification of DcPSY1 , DcCRITISO , DcD27 , DcCCD7 and Dc CCD8 transcript levels by real-time RT-PCR analysis normalized to equal levels of actin transcripts. All analyses were performed using three biological replicates. C: Control non-infested carrot roots; +: Carrot roots after P . aegyptiaca infestation. Means with the same letter are not significantly different from each other (Tukey’s test, P ≤ 0.05).
    Figure Legend Snippet: qRT-PCR determination of transcript levels of carrots ( D . carota ) DcPSY1 , DcCRTISO , DcD27 , DcCCD7 , and DcCCD8 in roots of different carrot cultivars before and after the infestion by obligate parasite P . aegyptiaca . Quantification of DcPSY1 , DcCRITISO , DcD27 , DcCCD7 and Dc CCD8 transcript levels by real-time RT-PCR analysis normalized to equal levels of actin transcripts. All analyses were performed using three biological replicates. C: Control non-infested carrot roots; +: Carrot roots after P . aegyptiaca infestation. Means with the same letter are not significantly different from each other (Tukey’s test, P ≤ 0.05).

    Techniques Used: Quantitative RT-PCR

    qRT-PCR determination of transcript levels of P . aegyptiaca PaD27 , PaCCD7 and PaCCD8 in the tubercles of the parasitic organs after infested carrot and tomato (as a control) roots. Quantification of PaD27 , PaCCD7 and PaCCD8 transcript level by real-time RT-PCR analysis normalized to equal levels of actin transcripts. All analyses were performed using three biological replicates. Means with the same letter are not significantly different from each other (Tukey’s test, P ≤ 0.05).
    Figure Legend Snippet: qRT-PCR determination of transcript levels of P . aegyptiaca PaD27 , PaCCD7 and PaCCD8 in the tubercles of the parasitic organs after infested carrot and tomato (as a control) roots. Quantification of PaD27 , PaCCD7 and PaCCD8 transcript level by real-time RT-PCR analysis normalized to equal levels of actin transcripts. All analyses were performed using three biological replicates. Means with the same letter are not significantly different from each other (Tukey’s test, P ≤ 0.05).

    Techniques Used: Quantitative RT-PCR

    22) Product Images from "AEG-1/miR-221 Axis Cooperatively Regulates the Progression of Hepatocellular Carcinoma by Targeting PTEN/PI3K/AKT Signaling Pathway"

    Article Title: AEG-1/miR-221 Axis Cooperatively Regulates the Progression of Hepatocellular Carcinoma by Targeting PTEN/PI3K/AKT Signaling Pathway

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20225526

    miR-221/AEG-1 regulates angiogenesis and cell cycle regulatory mRNA expressions in HCC cell line. Expression level of angiogenesis (LSF and MMP9) and cell cycle (p57, p53, and RB) regulatory mRNAs were analyzed by SYBER Green qRT-PCR in mock control, miR-mimic negative control, miR-inhibitor negative control, miR-221 mimic, miR-221 inhibitor, siRNA negative control, and AEG-1 siRNA transfected HepG2, Huh7, and Hep3B cells using GAPDH as an internal control. Error bars are presented as mean ± s.d. and * p
    Figure Legend Snippet: miR-221/AEG-1 regulates angiogenesis and cell cycle regulatory mRNA expressions in HCC cell line. Expression level of angiogenesis (LSF and MMP9) and cell cycle (p57, p53, and RB) regulatory mRNAs were analyzed by SYBER Green qRT-PCR in mock control, miR-mimic negative control, miR-inhibitor negative control, miR-221 mimic, miR-221 inhibitor, siRNA negative control, and AEG-1 siRNA transfected HepG2, Huh7, and Hep3B cells using GAPDH as an internal control. Error bars are presented as mean ± s.d. and * p

    Techniques Used: Expressing, Quantitative RT-PCR, Negative Control, Transfection

    23) Product Images from "Cytochrome c oxidase III as a mechanism for apoptosis in heart failure following myocardial infarction"

    Article Title: Cytochrome c oxidase III as a mechanism for apoptosis in heart failure following myocardial infarction

    Journal: American Journal of Physiology - Cell Physiology

    doi: 10.1152/ajpcell.00045.2009

    Overexpression and localization of COX III in mitochondria of HL-1 cells. A : agarose gel showing that the pEF-BOS-COX VIII-COX III-FLAG plasmid (pEF-BOS-C8C3F, 4 μg) was successfully transfected and overexpressed in HL-1 cells as detected by RT-PCR. Total length of the DNA sequence is 880 bp, which consists of 150 bp of COX VIII (mitochondrial targeted peptide), 780 bp of site-directed mutated COX III genes, and 30 bp of FLAG genes. MW, molecular weight. B : dose-dependent overexpression of COX III as a function of plasmid amount was detected by quantitative real-time PCR (qPCR). Different amounts of PEF-BOS-C8C3F plasmid (2, 4 and 6 μg) were transfected into 6-cm plates and cultured for 48 h. β-Actin was used as an internal control. Each condition was measured in triplicate. Values are means ± SE for the triplicates. * P
    Figure Legend Snippet: Overexpression and localization of COX III in mitochondria of HL-1 cells. A : agarose gel showing that the pEF-BOS-COX VIII-COX III-FLAG plasmid (pEF-BOS-C8C3F, 4 μg) was successfully transfected and overexpressed in HL-1 cells as detected by RT-PCR. Total length of the DNA sequence is 880 bp, which consists of 150 bp of COX VIII (mitochondrial targeted peptide), 780 bp of site-directed mutated COX III genes, and 30 bp of FLAG genes. MW, molecular weight. B : dose-dependent overexpression of COX III as a function of plasmid amount was detected by quantitative real-time PCR (qPCR). Different amounts of PEF-BOS-C8C3F plasmid (2, 4 and 6 μg) were transfected into 6-cm plates and cultured for 48 h. β-Actin was used as an internal control. Each condition was measured in triplicate. Values are means ± SE for the triplicates. * P

    Techniques Used: Over Expression, Agarose Gel Electrophoresis, Plasmid Preparation, Transfection, Reverse Transcription Polymerase Chain Reaction, Sequencing, Molecular Weight, Real-time Polymerase Chain Reaction, Cell Culture

    24) Product Images from "Transcriptional Regulation of BMP2 Expression by the PTH-CREB Signaling Pathway in Osteoblasts"

    Article Title: Transcriptional Regulation of BMP2 Expression by the PTH-CREB Signaling Pathway in Osteoblasts

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0020780

    PTH up-regulates BMP2 expression through CREB. (A, B) BMP2 mRNA levels in C2C12 cells, treated with PTH at 0 to 200 nM for 12 hours (A); or PTH at 50 nM for 0 to 12 hours (B), were measured by real time PCR with GAPDH normalization. * p
    Figure Legend Snippet: PTH up-regulates BMP2 expression through CREB. (A, B) BMP2 mRNA levels in C2C12 cells, treated with PTH at 0 to 200 nM for 12 hours (A); or PTH at 50 nM for 0 to 12 hours (B), were measured by real time PCR with GAPDH normalization. * p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    CREB activates BMP2 expression. (A) BMP2 mRNA levels in C2C12, 2T3 and UMR106 cells, transfected with CREB plasmid or vector for 24 hours, were measured by real time PCR with GAPDH106 cells. * p
    Figure Legend Snippet: CREB activates BMP2 expression. (A) BMP2 mRNA levels in C2C12, 2T3 and UMR106 cells, transfected with CREB plasmid or vector for 24 hours, were measured by real time PCR with GAPDH106 cells. * p

    Techniques Used: Expressing, Transfection, Plasmid Preparation, Real-time Polymerase Chain Reaction

    25) Product Images from "MicroRNA-574-3p, identified by microRNA library-based functional screening, modulates tamoxifen response in breast cancer"

    Article Title: MicroRNA-574-3p, identified by microRNA library-based functional screening, modulates tamoxifen response in breast cancer

    Journal: Scientific Reports

    doi: 10.1038/srep07641

    Identification of miR-574-3p target genes in breast cancer. (a) Schematic presentation of miR-574-3p target prediction by in silico analyses. Venn diagrams indicating numbers of candidate hits determined by 4 online prediction algorithms. Candidate genes commonly predicted by > 3 algorithms are described. (b) CLTC mRNA was upregulated in 4-hydroxytamoxifen (OHT)-resistant MCF-7 cells (OHTR cells). Expression levels of CLTC mRNA were determined by quantitative reverse transcription PCR (qRT-PCR) in parental MCF-7 cells and OHTR cells. Data are presented as mean ± SE in triplicate; ** P
    Figure Legend Snippet: Identification of miR-574-3p target genes in breast cancer. (a) Schematic presentation of miR-574-3p target prediction by in silico analyses. Venn diagrams indicating numbers of candidate hits determined by 4 online prediction algorithms. Candidate genes commonly predicted by > 3 algorithms are described. (b) CLTC mRNA was upregulated in 4-hydroxytamoxifen (OHT)-resistant MCF-7 cells (OHTR cells). Expression levels of CLTC mRNA were determined by quantitative reverse transcription PCR (qRT-PCR) in parental MCF-7 cells and OHTR cells. Data are presented as mean ± SE in triplicate; ** P

    Techniques Used: In Silico, Expressing, Polymerase Chain Reaction, Quantitative RT-PCR

    26) Product Images from "The polyphenol resveratrol promotes skeletal growth in mice through a sirtuin 1‐bone morphogenic protein 2 longevity axis) The polyphenol resveratrol promotes skeletal growth in mice through a sirtuin 1‐bone morphogenic protein 2 longevity axis"

    Article Title: The polyphenol resveratrol promotes skeletal growth in mice through a sirtuin 1‐bone morphogenic protein 2 longevity axis) The polyphenol resveratrol promotes skeletal growth in mice through a sirtuin 1‐bone morphogenic protein 2 longevity axis

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.14477

    eNOS and NO‐donors stimulate bone growth. eNOS mRNA expression in MC3T3 (A) and 2T3 (B) cells treated with RSV (1–100 μM) or vehicle for 24 h ( n = 5) determined by real time PCR. Nitrite (NO 2 − ) levels (C) or total eNOS protein (D) from 2T3 or MC3T3 cells treated with RSV (1–100 μM) or vehicle ( n = 5) for 48 h, determined by colorimetric assay. ALP levels from primary osteoblasts treated with NO‐donor (NOC22, 0.1–10 μM) or vehicle for 4 days ( n = 5) normalized to total cell protein (E). Calvariae from newborn mice cultured with NO‐donor (NOC22, 0.1–1 μM) or vehicle control for 4 days ( n = 5) processed for histology and H E staining (F). mRNA expression of osteoblast marker genes Runx2, osteocalcin (OCN) and BMP2 determined in 2T3 cells treated with NOC22 (1.0 μM) or vehicle for 24 h ( n = 5) by real time PCR (G). BMP2 protein levels assessed in conditioned media from NO‐donor‐treated osteoblasts (NOC22 or SNP, 3–200 μM, 48 h, n = 5), by ELISA (with rhBMP2 as standard) (H). The μCT analysis of dissected tibia from homozygous eNOS knockout ( −/− ) or wild‐type ( +/+ ) control mice (4 month, n = 10) (I) and trabecular bone volume (J) and BMD (K) analysis (* P
    Figure Legend Snippet: eNOS and NO‐donors stimulate bone growth. eNOS mRNA expression in MC3T3 (A) and 2T3 (B) cells treated with RSV (1–100 μM) or vehicle for 24 h ( n = 5) determined by real time PCR. Nitrite (NO 2 − ) levels (C) or total eNOS protein (D) from 2T3 or MC3T3 cells treated with RSV (1–100 μM) or vehicle ( n = 5) for 48 h, determined by colorimetric assay. ALP levels from primary osteoblasts treated with NO‐donor (NOC22, 0.1–10 μM) or vehicle for 4 days ( n = 5) normalized to total cell protein (E). Calvariae from newborn mice cultured with NO‐donor (NOC22, 0.1–1 μM) or vehicle control for 4 days ( n = 5) processed for histology and H E staining (F). mRNA expression of osteoblast marker genes Runx2, osteocalcin (OCN) and BMP2 determined in 2T3 cells treated with NOC22 (1.0 μM) or vehicle for 24 h ( n = 5) by real time PCR (G). BMP2 protein levels assessed in conditioned media from NO‐donor‐treated osteoblasts (NOC22 or SNP, 3–200 μM, 48 h, n = 5), by ELISA (with rhBMP2 as standard) (H). The μCT analysis of dissected tibia from homozygous eNOS knockout ( −/− ) or wild‐type ( +/+ ) control mice (4 month, n = 10) (I) and trabecular bone volume (J) and BMD (K) analysis (* P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Colorimetric Assay, ALP Assay, Mouse Assay, Cell Culture, Staining, Marker, Enzyme-linked Immunosorbent Assay, Knock-Out

    The eNOS‐SIRT1 axis is necessary for the pro‐osteogenic effects of RSV. Different effects on ALP levels (A) BMP2 gene expression (qPCR, B) and promoter activity (C) in primary osteoblasts from eNOS knockout ( −/− ) or wild‐type ( +/+ ) control mice ( n = 5) treated with RSV (5 μM) or vehicle (24 h). RSV‐induced ALP levels (5 μM) in the presence of the BMP inhibitor noggin (500 ng·mL −1 , 48 h, n = 5), normalized to total protein (D). SIRT1 gene expression in 2T3 osteoblasts following RSV (1–100 μM) treatment, quantified by real time PCR (E), along with eNOS (F) and BMP2 (G) mRNA levels transfected with SIRT1 siRNA or scramble control ( n = 5) with and without RSV treatment (5 μM). RSV‐induced ALP levels (5 μM) in the presence of SIRT1 (or scrambled control) siRNA (H) ( n = 5) (* P
    Figure Legend Snippet: The eNOS‐SIRT1 axis is necessary for the pro‐osteogenic effects of RSV. Different effects on ALP levels (A) BMP2 gene expression (qPCR, B) and promoter activity (C) in primary osteoblasts from eNOS knockout ( −/− ) or wild‐type ( +/+ ) control mice ( n = 5) treated with RSV (5 μM) or vehicle (24 h). RSV‐induced ALP levels (5 μM) in the presence of the BMP inhibitor noggin (500 ng·mL −1 , 48 h, n = 5), normalized to total protein (D). SIRT1 gene expression in 2T3 osteoblasts following RSV (1–100 μM) treatment, quantified by real time PCR (E), along with eNOS (F) and BMP2 (G) mRNA levels transfected with SIRT1 siRNA or scramble control ( n = 5) with and without RSV treatment (5 μM). RSV‐induced ALP levels (5 μM) in the presence of SIRT1 (or scrambled control) siRNA (H) ( n = 5) (* P

    Techniques Used: ALP Assay, Expressing, Real-time Polymerase Chain Reaction, Activity Assay, Knock-Out, Mouse Assay, Transfection

    27) Product Images from "Vacuolar ATPase ‘a2’ isoform exhibits distinct cell surface accumulation and modulates matrix metalloproteinase activity in ovarian cancer"

    Article Title: Vacuolar ATPase ‘a2’ isoform exhibits distinct cell surface accumulation and modulates matrix metalloproteinase activity in ovarian cancer

    Journal: Oncotarget

    doi:

    Expression profiling of V-ATPase ‘a’ subunit isoforms in ovarian cancer Real time PCR was used to quantify the relative mRNA amounts of V-ATPase V0‘a’ isoforms (V0a1/V0a3/V0a4) in ovarian cancer cell lines. The ovarian cancer cells exhibited higher mRNA levels of (A) V0a1 and (B) V0a3 isoforms in ovarian cancer cell lines compared to normal control epithelial cells of ovary. (C) V0a4 isoform levels were very low and variable in different cell lines. The Ct values were normalized against the Ct values obtained for GAPDH from the same preparation. The data are provided as Mean ± SD from 3 independent experiments. (*** p
    Figure Legend Snippet: Expression profiling of V-ATPase ‘a’ subunit isoforms in ovarian cancer Real time PCR was used to quantify the relative mRNA amounts of V-ATPase V0‘a’ isoforms (V0a1/V0a3/V0a4) in ovarian cancer cell lines. The ovarian cancer cells exhibited higher mRNA levels of (A) V0a1 and (B) V0a3 isoforms in ovarian cancer cell lines compared to normal control epithelial cells of ovary. (C) V0a4 isoform levels were very low and variable in different cell lines. The Ct values were normalized against the Ct values obtained for GAPDH from the same preparation. The data are provided as Mean ± SD from 3 independent experiments. (*** p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    Ovarian carcinoma cell lines exhibit high expression of V-ATPase-V0a2 on cell surface (A) Real time PCR analysis revealed higher relative mRNA levels of V-ATPase-V0a2 isoform in ovarian cancer cell lines compared to normal ovary epithelia. The Ct values were normalized against the Ct values for GAPDH from the same preparation. The data are provided as mean ± SD from 3 independent experiments. (*** p
    Figure Legend Snippet: Ovarian carcinoma cell lines exhibit high expression of V-ATPase-V0a2 on cell surface (A) Real time PCR analysis revealed higher relative mRNA levels of V-ATPase-V0a2 isoform in ovarian cancer cell lines compared to normal ovary epithelia. The Ct values were normalized against the Ct values for GAPDH from the same preparation. The data are provided as mean ± SD from 3 independent experiments. (*** p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    28) Product Images from "Green tea powder supplementation enhances fermentation and antioxidant activity of set-type yogurt"

    Article Title: Green tea powder supplementation enhances fermentation and antioxidant activity of set-type yogurt

    Journal: Food Science and Biotechnology

    doi: 10.1007/s10068-018-0370-9

    ( A ) Antioxidant effects of extracts of green tea powder (GTP) yogurt on human colorectal cells. Cells were pretreated with extracts of 0, 1, 2, or 3% GTP (w/v) yogurts for 15 h, before being stimulated with 1 µg/mL lipopolysaccharide (LPS) for 6 h, and stained with the fluorogenic dye DCFDA to detect H 2 O 2 production. The intensity of green fluorescence was assessed using a fluorescence microscope and analyzed with ImageJ. The images shown here are representative of three independent experiments. ( B ) Western blot analysis of the antioxidant proteins Nrf2 and HO-1 in HT-29 cells. Protein expression in cells treated for 15 h with yogurt extract supplemented with 0, 1, 2, or 3% GTP (w/v) was measured. GAPDH levels were measured as a loading control. The blot shown is representative of three independent experiments. Protein bands were quantified using ImageJ software. ( C ) mRNA expression of IL-1β and TNF-α in HT-29 cells. Cells were pretreated with extracts of 0, 1, 2, or 3% GTP (w/v) yogurts for 15 h, followed by stimulation with 1 µg/mL LPS for 12 h. The mRNA expression levels were measured using quantitative real-time PCR technique. GAPDH was used as a control gene. In all graphs, values are mean ± SEM (n = 3). *Significant differences versus control ( p
    Figure Legend Snippet: ( A ) Antioxidant effects of extracts of green tea powder (GTP) yogurt on human colorectal cells. Cells were pretreated with extracts of 0, 1, 2, or 3% GTP (w/v) yogurts for 15 h, before being stimulated with 1 µg/mL lipopolysaccharide (LPS) for 6 h, and stained with the fluorogenic dye DCFDA to detect H 2 O 2 production. The intensity of green fluorescence was assessed using a fluorescence microscope and analyzed with ImageJ. The images shown here are representative of three independent experiments. ( B ) Western blot analysis of the antioxidant proteins Nrf2 and HO-1 in HT-29 cells. Protein expression in cells treated for 15 h with yogurt extract supplemented with 0, 1, 2, or 3% GTP (w/v) was measured. GAPDH levels were measured as a loading control. The blot shown is representative of three independent experiments. Protein bands were quantified using ImageJ software. ( C ) mRNA expression of IL-1β and TNF-α in HT-29 cells. Cells were pretreated with extracts of 0, 1, 2, or 3% GTP (w/v) yogurts for 15 h, followed by stimulation with 1 µg/mL LPS for 12 h. The mRNA expression levels were measured using quantitative real-time PCR technique. GAPDH was used as a control gene. In all graphs, values are mean ± SEM (n = 3). *Significant differences versus control ( p

    Techniques Used: Staining, Fluorescence, Microscopy, Western Blot, Expressing, Software, Real-time Polymerase Chain Reaction

    29) Product Images from "miR-92b controls glioma proliferation and invasion through regulating Wnt/beta-catenin signaling via Nemo-like kinase"

    Article Title: miR-92b controls glioma proliferation and invasion through regulating Wnt/beta-catenin signaling via Nemo-like kinase

    Journal: Neuro-Oncology

    doi: 10.1093/neuonc/not004

    miR-92b I suppresses glioma cell proliferation and invasion. (A) miR-92b expression was quantified by qRT-PCR analysis. miR-92b inhibitor significantly suppressed miR-92b expression in both SNB19 and LN229 cells ( P
    Figure Legend Snippet: miR-92b I suppresses glioma cell proliferation and invasion. (A) miR-92b expression was quantified by qRT-PCR analysis. miR-92b inhibitor significantly suppressed miR-92b expression in both SNB19 and LN229 cells ( P

    Techniques Used: Expressing, Quantitative RT-PCR

    miR-92b is upregulated in glioma samples and glioma cells. (A) miR-92b expression in different grade glioma and normal brain tissues by qRT-PCR. (B) qRT-PCR analysis showed that U251, U87, LN229, SNB19, and A172 glioma cells express high levels of miR-92b,
    Figure Legend Snippet: miR-92b is upregulated in glioma samples and glioma cells. (A) miR-92b expression in different grade glioma and normal brain tissues by qRT-PCR. (B) qRT-PCR analysis showed that U251, U87, LN229, SNB19, and A172 glioma cells express high levels of miR-92b,

    Techniques Used: Expressing, Quantitative RT-PCR

    30) Product Images from "Paraventricular Nucleus Sim1 Neuron Ablation Mediated Obesity Is Resistant to High Fat Diet"

    Article Title: Paraventricular Nucleus Sim1 Neuron Ablation Mediated Obesity Is Resistant to High Fat Diet

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0081087

    Real-time quantitative PCR comparing Sim1 mRNA, MC4R mRNA, and Galanin mRNA in PVN and Amygdala of Sim1creiDTR and iDTR mice, and comparing OXTmRNA in PVN of Sim1creiDTR and iDTR mice. Relative mRNA abundance of SIM1 (A), MC4R (B), OXT (C), and GAL (F) expression in the PVN and SIM1 (D), MC4R (E), and GAL (G) amygdala. (n=4 to 9 for each group, * p
    Figure Legend Snippet: Real-time quantitative PCR comparing Sim1 mRNA, MC4R mRNA, and Galanin mRNA in PVN and Amygdala of Sim1creiDTR and iDTR mice, and comparing OXTmRNA in PVN of Sim1creiDTR and iDTR mice. Relative mRNA abundance of SIM1 (A), MC4R (B), OXT (C), and GAL (F) expression in the PVN and SIM1 (D), MC4R (E), and GAL (G) amygdala. (n=4 to 9 for each group, * p

    Techniques Used: Real-time Polymerase Chain Reaction, Mouse Assay, Expressing

    31) Product Images from "Identification, characterization and expression analysis of passion fruit (Passiflora edulis) microRNAs"

    Article Title: Identification, characterization and expression analysis of passion fruit (Passiflora edulis) microRNAs

    Journal: 3 Biotech

    doi: 10.1007/s13205-019-2000-5

    Validation of selected passion fruit miRNAs (ped-miR160, ped-miR164, ped-miR166, ped-miR393, ped-miR394, and ped-miR398) by semiquantitative reverse transcription PCR (fruit tissues). The resulting PCR products were checked in 2% agarose gel with EtBr staining. U6 was employed as a positive control
    Figure Legend Snippet: Validation of selected passion fruit miRNAs (ped-miR160, ped-miR164, ped-miR166, ped-miR393, ped-miR394, and ped-miR398) by semiquantitative reverse transcription PCR (fruit tissues). The resulting PCR products were checked in 2% agarose gel with EtBr staining. U6 was employed as a positive control

    Techniques Used: Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining, Positive Control

    32) Product Images from "RNA sequencing reveals resistance of TLR4 ligand-activated microglial cells to inflammation mediated by the selective jumonji H3K27 demethylase inhibitor"

    Article Title: RNA sequencing reveals resistance of TLR4 ligand-activated microglial cells to inflammation mediated by the selective jumonji H3K27 demethylase inhibitor

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-06914-5

    In vivo effect of GSK-J4 on pro-inflammatory responses in LPS-challenged mice. ICR mice (n = 5 for each group) were treated with LPS (1 mg/kg) following GSK-J4 injection (1 mg/kg) and brain microglia were collected at 4-h to determine LPS-induced gene expression by qRT-PCR. The CCL2, CCL3, CCL4, CCL12, IRF1, IL1A, IL1B, and IRG1 genes were significantly down-regulated in the GSK-J4-injected adult microglial cells. Gene expression was normalized to GAPDH transcript levels. Each point represents data from an individual mouse, all values shown as mean ± S.E.M. * P
    Figure Legend Snippet: In vivo effect of GSK-J4 on pro-inflammatory responses in LPS-challenged mice. ICR mice (n = 5 for each group) were treated with LPS (1 mg/kg) following GSK-J4 injection (1 mg/kg) and brain microglia were collected at 4-h to determine LPS-induced gene expression by qRT-PCR. The CCL2, CCL3, CCL4, CCL12, IRF1, IL1A, IL1B, and IRG1 genes were significantly down-regulated in the GSK-J4-injected adult microglial cells. Gene expression was normalized to GAPDH transcript levels. Each point represents data from an individual mouse, all values shown as mean ± S.E.M. * P

    Techniques Used: In Vivo, Mouse Assay, Injection, Expressing, Quantitative RT-PCR

    33) Product Images from "Transcriptional control of DNA replication licensing by Myc"

    Article Title: Transcriptional control of DNA replication licensing by Myc

    Journal: Scientific Reports

    doi: 10.1038/srep03444

    In vivo binding of Myc to the quail and chicken CDT1 promoters. (a) ChIP analysis using chromatin from normal QEFs, or from QEF/MC29 and QEF/Rc-Myc transformed by the v-Myc protein. (b) ChIP using chromatin from normal CEF and from CEF/MC29. (c) ChIP using chromatin from normal QEF, from the chemically transformed quail cell line QT6, or from the v- myc -transformed cell lines QEF/MC29, QEF/Rc-Myc, and Q8. The QEF(CEF)/MC29 cells and the Q8 cell line are transformed by the p110 Gag-Myc hybrid protein, whereas the QEF/Rc-Myc cells are transformed by a v-Myc protein not fused to viral structural protein sequences. Antisera directed against Myc, Max, or Gag were used for immunoprecipitation, followed by PCR amplification of DNA from the quail CDT1 (241 bp fragment) or chicken CDT1 (253 bp fragment) regulatory regions both containing two canonical Myc binding sites, or of DNA from the quail WS5 (290 or 273 bp fragment) and chicken WS5 ( Mmp115 ) (293 bp fragment) promoters both containing four Myc binding sites 33 . Primers specific for a chicken BASP1 exon 2 fragment (197 bp) containing no E-boxes (control), and reactions with normal rabbit serum (NRS) or 1.6% of total chromatin (input) were used as controls. Fragments were analyzed by agarose (1.5% wt/vol) gel electrophoresis. (d) ChIP-quantitative PCR (ChIP-qPCR) using chromatin from normal QEF, or from QEF/MC29 and QEF/Rc-Myc. Immunoprecipitated DNA was analyzed in triplicate by qPCR using primers to amplify a 167-bp segment of the regulatory region from the quail CDT1 promoter containing the two canonical Myc binding sites. Signals were normalized to input chromatin and shown as % input. Full-length gels are included in the supplementary information .
    Figure Legend Snippet: In vivo binding of Myc to the quail and chicken CDT1 promoters. (a) ChIP analysis using chromatin from normal QEFs, or from QEF/MC29 and QEF/Rc-Myc transformed by the v-Myc protein. (b) ChIP using chromatin from normal CEF and from CEF/MC29. (c) ChIP using chromatin from normal QEF, from the chemically transformed quail cell line QT6, or from the v- myc -transformed cell lines QEF/MC29, QEF/Rc-Myc, and Q8. The QEF(CEF)/MC29 cells and the Q8 cell line are transformed by the p110 Gag-Myc hybrid protein, whereas the QEF/Rc-Myc cells are transformed by a v-Myc protein not fused to viral structural protein sequences. Antisera directed against Myc, Max, or Gag were used for immunoprecipitation, followed by PCR amplification of DNA from the quail CDT1 (241 bp fragment) or chicken CDT1 (253 bp fragment) regulatory regions both containing two canonical Myc binding sites, or of DNA from the quail WS5 (290 or 273 bp fragment) and chicken WS5 ( Mmp115 ) (293 bp fragment) promoters both containing four Myc binding sites 33 . Primers specific for a chicken BASP1 exon 2 fragment (197 bp) containing no E-boxes (control), and reactions with normal rabbit serum (NRS) or 1.6% of total chromatin (input) were used as controls. Fragments were analyzed by agarose (1.5% wt/vol) gel electrophoresis. (d) ChIP-quantitative PCR (ChIP-qPCR) using chromatin from normal QEF, or from QEF/MC29 and QEF/Rc-Myc. Immunoprecipitated DNA was analyzed in triplicate by qPCR using primers to amplify a 167-bp segment of the regulatory region from the quail CDT1 promoter containing the two canonical Myc binding sites. Signals were normalized to input chromatin and shown as % input. Full-length gels are included in the supplementary information .

    Techniques Used: In Vivo, Binding Assay, Chromatin Immunoprecipitation, Transformation Assay, Immunoprecipitation, Polymerase Chain Reaction, Amplification, Nucleic Acid Electrophoresis, Real-time Polymerase Chain Reaction

    34) Product Images from "Sulforaphane reactivates cellular antioxidant defense by inducing Nrf2/ARE/Prdx6 activity during aging and oxidative stress"

    Article Title: Sulforaphane reactivates cellular antioxidant defense by inducing Nrf2/ARE/Prdx6 activity during aging and oxidative stress

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-14520-8

    In vivo DNA binding assay revealed that SFN reinforced binding activity of Nrf2 in SRA-hLECs and aging/aged primary hLECs. ( A ) Schematic representation of the regulatory region of proximal promoter of human Prdx6 gene-containing ARE binding sites showing primer location and sequences used in ChIP assay. (B) SFN induced increase in DNA binding activity of Nrf2 to Prdx6 gene promoter containing ARE site in SRA-hLECs. ChIP experiment was carried out by using ChIP-IT® Express and ChIP-IT® qPCR analysis Kit. Chromatin samples prepared from SRA-hLECs treated with varying concentrations (0, 3 µM and 6 µM) of SFN for 24 h were subjected to ChIP assay with a ChIP grade antibody, anti-Nrf2 (black bars) and control IgG (gray bars). The DNA fragments were used as templates for qPCR by using primers designed to amplify −400 to −305 region of the human Prdx6 promoter bearing Nrf2/ARE sites as shown. Histogram shows the amplified DNA band visualized with real-time PCR analysis. DMSO (0) vs 3 µM and 6 µM SFN and 3 µM vs 6 µM SFN treatment; *p
    Figure Legend Snippet: In vivo DNA binding assay revealed that SFN reinforced binding activity of Nrf2 in SRA-hLECs and aging/aged primary hLECs. ( A ) Schematic representation of the regulatory region of proximal promoter of human Prdx6 gene-containing ARE binding sites showing primer location and sequences used in ChIP assay. (B) SFN induced increase in DNA binding activity of Nrf2 to Prdx6 gene promoter containing ARE site in SRA-hLECs. ChIP experiment was carried out by using ChIP-IT® Express and ChIP-IT® qPCR analysis Kit. Chromatin samples prepared from SRA-hLECs treated with varying concentrations (0, 3 µM and 6 µM) of SFN for 24 h were subjected to ChIP assay with a ChIP grade antibody, anti-Nrf2 (black bars) and control IgG (gray bars). The DNA fragments were used as templates for qPCR by using primers designed to amplify −400 to −305 region of the human Prdx6 promoter bearing Nrf2/ARE sites as shown. Histogram shows the amplified DNA band visualized with real-time PCR analysis. DMSO (0) vs 3 µM and 6 µM SFN and 3 µM vs 6 µM SFN treatment; *p

    Techniques Used: In Vivo, DNA Binding Assay, Binding Assay, Activity Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Amplification

    SFN induced Nrf2 expression and enhanced nuclear accumulation in both SRA-hLECs and rLECs. ( A ) Effect of SFN concentration(s) on expression of Nrf2 mRNA in SRA-hLECs. Total RNA was isolated and real-time PCR was performed using specific primers. mRNA expression of Nrf2 was adjusted/normalized to the mRNA copies of β-actin. Histogram represents mean ± S.D. obtained from three independent experiments. Open vs gray and black bars; gray vs black bar; * p
    Figure Legend Snippet: SFN induced Nrf2 expression and enhanced nuclear accumulation in both SRA-hLECs and rLECs. ( A ) Effect of SFN concentration(s) on expression of Nrf2 mRNA in SRA-hLECs. Total RNA was isolated and real-time PCR was performed using specific primers. mRNA expression of Nrf2 was adjusted/normalized to the mRNA copies of β-actin. Histogram represents mean ± S.D. obtained from three independent experiments. Open vs gray and black bars; gray vs black bar; * p

    Techniques Used: Expressing, Concentration Assay, Isolation, Real-time Polymerase Chain Reaction

    35) Product Images from "Real-time PCR for direct aptamer quantification on functionalized graphene surfaces"

    Article Title: Real-time PCR for direct aptamer quantification on functionalized graphene surfaces

    Journal: Scientific Reports

    doi: 10.1038/s41598-019-55892-3

    Representative results obtained using qPCR primer sets in conventional and qPCR. (a) 2% typical agarose gel stained with ethidium bromide showing the amplification of the sequence of approximately 56 bp, referent of SA20 aptamer at the end of the PCR process. Molecular weight (M.W.), negative template control (NTC, lane 2), positive control (lane 3) and a SA20-pyrene functionalized graphene (lane 4). Full-length unmodified gel is presented in Supplementary Figure 1. (b) Amplification curves of real-time PCR (using 10-fold serial dilutions of SA20 aptamer ranging from 0.5 ng to 0.00005 ng) representing the amplification of each point in the standard curve and the amplification of the positive control (with 0.5 ng of SA20). (c) Standard curve in which the Ct values were plotted against the template quantity. These curves ( b and c ) are representative of 10 independent experiments, all presenting similar results.
    Figure Legend Snippet: Representative results obtained using qPCR primer sets in conventional and qPCR. (a) 2% typical agarose gel stained with ethidium bromide showing the amplification of the sequence of approximately 56 bp, referent of SA20 aptamer at the end of the PCR process. Molecular weight (M.W.), negative template control (NTC, lane 2), positive control (lane 3) and a SA20-pyrene functionalized graphene (lane 4). Full-length unmodified gel is presented in Supplementary Figure 1. (b) Amplification curves of real-time PCR (using 10-fold serial dilutions of SA20 aptamer ranging from 0.5 ng to 0.00005 ng) representing the amplification of each point in the standard curve and the amplification of the positive control (with 0.5 ng of SA20). (c) Standard curve in which the Ct values were plotted against the template quantity. These curves ( b and c ) are representative of 10 independent experiments, all presenting similar results.

    Techniques Used: Real-time Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining, Amplification, Sequencing, Polymerase Chain Reaction, Molecular Weight, Positive Control

    36) Product Images from "miR-199a-3p negatively regulates the progression of osteosarcoma through targeting AXL"

    Article Title: miR-199a-3p negatively regulates the progression of osteosarcoma through targeting AXL

    Journal: American Journal of Cancer Research

    doi:

    A. The relative expression of miR-199a-3p was detected by qRT-PCR in 33 cases of paraffin-embedded osteosarcoma tissues and paired normal muscle tissues. The expression of miR-199a-3p was clearly lower in tumors than in paired normal tissues; B. In those
    Figure Legend Snippet: A. The relative expression of miR-199a-3p was detected by qRT-PCR in 33 cases of paraffin-embedded osteosarcoma tissues and paired normal muscle tissues. The expression of miR-199a-3p was clearly lower in tumors than in paired normal tissues; B. In those

    Techniques Used: Expressing, Quantitative RT-PCR

    37) Product Images from "Circular RNA DLGAP4 Ameliorates Ischemic Stroke Outcomes by Targeting miR-143 to Regulate Endothelial-Mesenchymal Transition Associated with Blood–Brain Barrier Integrity"

    Article Title: Circular RNA DLGAP4 Ameliorates Ischemic Stroke Outcomes by Targeting miR-143 to Regulate Endothelial-Mesenchymal Transition Associated with Blood–Brain Barrier Integrity

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.1348-17.2017

    circDLGAP4 binds miR-143 and is downregulated in the plasma of AIS patients and tMCAO mice. A , Left, circDLGAP4 contains one site that is complementary to miR-143 according to the bioinformatics program RNA hybrid. Right, A biotin-coupled miR-143 mutant. B , Divergent primers amplified circDLGAP4 from cDNA, but not genomic DNA (gDNA). GAPDH, linear control; M, marker. C , Examination of circDLGAP4 levels in the plasma of healthy controls and AIS patients via RT-PCR. circDLGAP4 levels were decreased in AIS patients by 0.6-fold the levels observed in healthy controls. n = 26 individuals/group. *** p
    Figure Legend Snippet: circDLGAP4 binds miR-143 and is downregulated in the plasma of AIS patients and tMCAO mice. A , Left, circDLGAP4 contains one site that is complementary to miR-143 according to the bioinformatics program RNA hybrid. Right, A biotin-coupled miR-143 mutant. B , Divergent primers amplified circDLGAP4 from cDNA, but not genomic DNA (gDNA). GAPDH, linear control; M, marker. C , Examination of circDLGAP4 levels in the plasma of healthy controls and AIS patients via RT-PCR. circDLGAP4 levels were decreased in AIS patients by 0.6-fold the levels observed in healthy controls. n = 26 individuals/group. *** p

    Techniques Used: Mouse Assay, Mutagenesis, Amplification, Marker, Reverse Transcription Polymerase Chain Reaction

    circDLGAP4 inhibits EndoMT targeting of miR-143 in mouse brain endothelial cells. A , Transduction of circDLGAP4 lentiviruses increased circDLGAP4 expression in mouse brain endothelial bEnd.3 cells, as determined via RT-PCR. Data are mean ± SEM of three independent experiments. ** p = 0.002, circDLGAP4 versus circControl (Student's t test). B , Effects of circDLGAP4 overexpression on TJP expression in mouse brain endothelial bEnd.3 cells treated with OGD/R. Transduction of cells with circDLGAP4 lentiviruses significantly inhibited the decrease in the expression of TJP induced by OGD/R. Claudin-5, F (3,8) = 41.632: ** p = 0.002, circControl OGD/R versus circControl Control. ## p = 0.002, circDLGAP4 OGD/R versus circControl OGD/R. Occludin, F (3,8) = 30.396: * p = 0.031, circControl OGD/R versus circControl Control. ## p = 0.003, circDLGAP4 OGD/R versus circControl OGD/R. ZO-1, F (3,8) = 24.985: ** p = 0.009, circControl OGD/R versus circControl Control. ## p = 0.004, circDLGAP4 OGD/R versus circControl OGD/R (one-way ANOVA followed by the Holm–Sidak Test). C , Effects of circDLGAP4 overexpression on mesenchymal cell marker expression in mouse brain endothelial bEnd.3 cells treated with OGD/R. Transduction of cells with circDLGAP4 lentiviruses significantly inhibited the increased expression of mesenchymal cell markers induced by OGD/R. Col I, F (3,8) = 32.075: *** p
    Figure Legend Snippet: circDLGAP4 inhibits EndoMT targeting of miR-143 in mouse brain endothelial cells. A , Transduction of circDLGAP4 lentiviruses increased circDLGAP4 expression in mouse brain endothelial bEnd.3 cells, as determined via RT-PCR. Data are mean ± SEM of three independent experiments. ** p = 0.002, circDLGAP4 versus circControl (Student's t test). B , Effects of circDLGAP4 overexpression on TJP expression in mouse brain endothelial bEnd.3 cells treated with OGD/R. Transduction of cells with circDLGAP4 lentiviruses significantly inhibited the decrease in the expression of TJP induced by OGD/R. Claudin-5, F (3,8) = 41.632: ** p = 0.002, circControl OGD/R versus circControl Control. ## p = 0.002, circDLGAP4 OGD/R versus circControl OGD/R. Occludin, F (3,8) = 30.396: * p = 0.031, circControl OGD/R versus circControl Control. ## p = 0.003, circDLGAP4 OGD/R versus circControl OGD/R. ZO-1, F (3,8) = 24.985: ** p = 0.009, circControl OGD/R versus circControl Control. ## p = 0.004, circDLGAP4 OGD/R versus circControl OGD/R (one-way ANOVA followed by the Holm–Sidak Test). C , Effects of circDLGAP4 overexpression on mesenchymal cell marker expression in mouse brain endothelial bEnd.3 cells treated with OGD/R. Transduction of cells with circDLGAP4 lentiviruses significantly inhibited the increased expression of mesenchymal cell markers induced by OGD/R. Col I, F (3,8) = 32.075: *** p

    Techniques Used: Transduction, Expressing, Reverse Transcription Polymerase Chain Reaction, Over Expression, Marker

    38) Product Images from "G?12 Drives Invasion of Oral Squamous Cell Carcinoma through Up-Regulation of Proinflammatory Cytokines"

    Article Title: G?12 Drives Invasion of Oral Squamous Cell Carcinoma through Up-Regulation of Proinflammatory Cytokines

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0066133

    Gα 12 -dependent regulation of IL-6 and IL-8 in OSCC. (A) Dot plots of the linear regression analysis showing a positive correlation of gene expressions between Gα 12 and IL-6/IL-8 in OSCC tumors and normal mucosa tissues. The relative expression scales are shown by RMA value in the microarray data. (B) IL-6 and IL-8 mRNA levels are positively regulated by Gα 12 in OSCC cells. Gα 12 levels in four different OSCC cell lines (HSC-3, SCC25, OC3, and CGHNC9) were altered by the transient overexpression or RNAi knockdown of Gα 12 . The qPCR results were normalized against GAPDH. The lower panel shows the electrophoresis image of the RT-PCR products. (C) The secreted proteins of IL-6 and IL-8 are up-regulated by Gα 12 in OSCC cells. ELISA assay was used to measure IL-6 and IL-8 in the conditioned media of the Gα 12 -overexpressing or -depleted HSC-3, SCC25, OC-3, and CGHNC9 cells. (D) The LPA-induced IL-6 and IL-8 production is regulated by Gα 12 . IL-6 and IL-8 protein levels in conditioned media of OSCC cells were measured by ELISA assay. The baseline IL-6 and IL-8 levels in conditioned media from four different OSCC cell lines are shown in Figure S2 . All the quantitative results are expressed as a fold change relative to the controls. The statistical results were analyzed by t-test , * P
    Figure Legend Snippet: Gα 12 -dependent regulation of IL-6 and IL-8 in OSCC. (A) Dot plots of the linear regression analysis showing a positive correlation of gene expressions between Gα 12 and IL-6/IL-8 in OSCC tumors and normal mucosa tissues. The relative expression scales are shown by RMA value in the microarray data. (B) IL-6 and IL-8 mRNA levels are positively regulated by Gα 12 in OSCC cells. Gα 12 levels in four different OSCC cell lines (HSC-3, SCC25, OC3, and CGHNC9) were altered by the transient overexpression or RNAi knockdown of Gα 12 . The qPCR results were normalized against GAPDH. The lower panel shows the electrophoresis image of the RT-PCR products. (C) The secreted proteins of IL-6 and IL-8 are up-regulated by Gα 12 in OSCC cells. ELISA assay was used to measure IL-6 and IL-8 in the conditioned media of the Gα 12 -overexpressing or -depleted HSC-3, SCC25, OC-3, and CGHNC9 cells. (D) The LPA-induced IL-6 and IL-8 production is regulated by Gα 12 . IL-6 and IL-8 protein levels in conditioned media of OSCC cells were measured by ELISA assay. The baseline IL-6 and IL-8 levels in conditioned media from four different OSCC cell lines are shown in Figure S2 . All the quantitative results are expressed as a fold change relative to the controls. The statistical results were analyzed by t-test , * P

    Techniques Used: Expressing, Microarray, Over Expression, Real-time Polymerase Chain Reaction, Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    39) Product Images from "Anthocyanin-rich fractions from red raspberries attenuate inflammation in both RAW264.7 macrophages and a mouse model of colitis"

    Article Title: Anthocyanin-rich fractions from red raspberries attenuate inflammation in both RAW264.7 macrophages and a mouse model of colitis

    Journal: Scientific Reports

    doi: 10.1038/srep06234

    RR-ARFs inhibit LPS/IFN-γ-mediated iNOS, COX-2, IL-1β and IL-6 expressions in RAW264.7 cells. Cells were incubated with different extracts of RR and LPS/IFN-γ for 16 h respectively at concentrations indicated. The expressions of mRNA for iNOS (a), COX-2 (b), IL-1β (c) and IL-6 (d) were analyzed by real-time PCR normalized to RPL14 mRNA. iNOS (130 kDa) and COX-2 (73 kDa) protein levels (e) were examined using Western blot. β-actin was used as an internal loading control. The full-length blots with anti-iNOS, anti-COX-2 and anti-β-actin antibodies were presented in supplementary Figure S1 . All gels have been run simultaneously under the same experimental conditions. Cell culture experiments were performed at least three times. The relative fold changes of iNOS and COX-2 protein were compared with LPS/IFN-γ control (f). Values are expressed as the means ± SD of at least three independent experiments. * P
    Figure Legend Snippet: RR-ARFs inhibit LPS/IFN-γ-mediated iNOS, COX-2, IL-1β and IL-6 expressions in RAW264.7 cells. Cells were incubated with different extracts of RR and LPS/IFN-γ for 16 h respectively at concentrations indicated. The expressions of mRNA for iNOS (a), COX-2 (b), IL-1β (c) and IL-6 (d) were analyzed by real-time PCR normalized to RPL14 mRNA. iNOS (130 kDa) and COX-2 (73 kDa) protein levels (e) were examined using Western blot. β-actin was used as an internal loading control. The full-length blots with anti-iNOS, anti-COX-2 and anti-β-actin antibodies were presented in supplementary Figure S1 . All gels have been run simultaneously under the same experimental conditions. Cell culture experiments were performed at least three times. The relative fold changes of iNOS and COX-2 protein were compared with LPS/IFN-γ control (f). Values are expressed as the means ± SD of at least three independent experiments. * P

    Techniques Used: Incubation, Real-time Polymerase Chain Reaction, Western Blot, Cell Culture

    40) Product Images from "FOXK1 promotes cell growth through activating wnt/β-catenin pathway and emerges as a novel target of miR-137 in glioma"

    Article Title: FOXK1 promotes cell growth through activating wnt/β-catenin pathway and emerges as a novel target of miR-137 in glioma

    Journal: American Journal of Translational Research

    doi:

    FOXK1 is upregulated in glioma tissues and cell lines. A. The mRNA levels of FOXK1 in glioma tissues (Tumor) and normal brain tissues (Normal) as determined by qRT-PCR. B. Representative images of FOXK1 protein expression in glioma tissues (T) and normal brain tissues (N) as detected by western blot. C. The mRNA levels of FOXK1 in glioma cell lines A172, U251, Ln229, U87 and normal human astrocytes (NHA) as determined by qRT-PCR. D. The FOXK1 protein expression in four glioma cell lines and normal human astrocytes (NHA) as detected by western blot. Data were mean ± SD. *P
    Figure Legend Snippet: FOXK1 is upregulated in glioma tissues and cell lines. A. The mRNA levels of FOXK1 in glioma tissues (Tumor) and normal brain tissues (Normal) as determined by qRT-PCR. B. Representative images of FOXK1 protein expression in glioma tissues (T) and normal brain tissues (N) as detected by western blot. C. The mRNA levels of FOXK1 in glioma cell lines A172, U251, Ln229, U87 and normal human astrocytes (NHA) as determined by qRT-PCR. D. The FOXK1 protein expression in four glioma cell lines and normal human astrocytes (NHA) as detected by western blot. Data were mean ± SD. *P

    Techniques Used: Quantitative RT-PCR, Expressing, Western Blot

    FOXK1 overexpression abolished the suppressive effect of miR-137 in glioma. A. Glioma cell lines U251 and Ln229 were transfected with miR-137 mimics or miR-NC. The relative expression levels of miR-137 were detected by qRT-PCR. B and C. U251 or Ln229 was transfected with miR-137 or/and FOXK1 plasmid to restore the expression of FOXK1 as indicated. Cell proliferation was determined by CCK-8 assay. D and E. Apoptosis rates of transfected U251 or Ln229 cells as detected by flow cytometry. F. The inverse correlation between relative FOXK1 mRNA levels and relative miR-137 expression in 30 glioma tissue samples. Data were mean ± SD. *P
    Figure Legend Snippet: FOXK1 overexpression abolished the suppressive effect of miR-137 in glioma. A. Glioma cell lines U251 and Ln229 were transfected with miR-137 mimics or miR-NC. The relative expression levels of miR-137 were detected by qRT-PCR. B and C. U251 or Ln229 was transfected with miR-137 or/and FOXK1 plasmid to restore the expression of FOXK1 as indicated. Cell proliferation was determined by CCK-8 assay. D and E. Apoptosis rates of transfected U251 or Ln229 cells as detected by flow cytometry. F. The inverse correlation between relative FOXK1 mRNA levels and relative miR-137 expression in 30 glioma tissue samples. Data were mean ± SD. *P

    Techniques Used: Over Expression, Transfection, Expressing, Quantitative RT-PCR, Plasmid Preparation, CCK-8 Assay, Flow Cytometry, Cytometry

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    Thermo Fisher quantitative real time reverse transcription polymerase chain reaction qrt pcr
    Knockdown or suppression of p65 reduces GLS1 expression in HCC cells. ( A ) Western blot shows the knockdown of c-Myc without interfering with GLS1 expression in HepG2 and Hep3B cells. ( B ) Western blot shows that p65 knockdown reduces GLS1 protein levels in both HepG2 and Hep3B cells. ( C ) <t>qRT-PCR</t> indicates that p65 knockdown reduces GLS1 mRNA levels in both HepG2 and Hep3B cells. ** p
    Quantitative Real Time Reverse Transcription Polymerase Chain Reaction Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher quantitative real time pcr qrt pcr
    LINC01296 knockdown suppressed the migration of bladder cancer cells. Notes: ( A , B ) Transwell assays were used to measure the migration ability of T24 and 5637 cells transfected with si-LINC01296 or si-NC. ( C , D ) Wound healing assays were used to measure the migration ability of T24 and 5637 cells transfected with si-LINC01296 or si-NC. ( E ) <t>qRT-</t> <t>PCR</t> analyses were used to measure mRNA levels of E-cadherin and N-cadherin following the treatment of bladder cells with anti-LINC01296 siRNA. ( F ) Western blot assays were used to measure protein levels of E-cadherin and N-cadherin following the treatment of bladder cells with anti-LINC01296 siRNA. Data are presented as mean ± SD of three independent experiments. *P
    Quantitative Real Time Pcr Qrt Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1152 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher quantitative real time polymerase chain reaction qrt pcr total rna
    Up-regulation of miR-96 and down-regulation of FOXO1 in tumor tissues and HepG2 cells. a miR-96 relative expression in tumor tissues and non-tumor tissues by <t>qRT-PCR;</t> b FOXO1 mRNA relative expression in tumor tissues and non-tumor tissues by qRT-PCR; c FOXO1 protein relative expression in tumor tissues and non-tumor tissues by Western blot; d miR-96 relative expression in L02 and HepG2 cells by qRT-PCR; e FOXO1 mRNA relative expression in L02 and HepG2 cells by qRT-PCR; f FOXO1 protein relative expression in L02 and HepG2 cells by Western blot. ** P
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    Thermo Fisher real time qrt pcr frozen day 60 liver
    Serum AAT c-myc Expression, Muscle Immunohistochemistry for AAT, and <t>RT-PCR</t> (A) Western blot gel image for all time points (please note that several AAV8 <t>day-60</t> samples were missing). (B) Semi-quantitation of western blot (WB) data represented as mean ± SEM. (C) Immunohistochemistry for AAT at day-60 representative sections for each dosing route for both AAV1 and AAV8. Muscle samples were collected for immunohistochemistry (IHC) from the level of the IM injections in both the quadriceps and gastrocnemius. (D) RT-PCR data from day-60 muscle and liver tissue. The gastrocnemius muscle samples were used for all groups. IM muscle samples were collected from the injection site. IM, intramuscular; IAPD, intra-arterial push and dwell; VLP, venous limb perfusion.
    Real Time Qrt Pcr Frozen Day 60 Liver, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Knockdown or suppression of p65 reduces GLS1 expression in HCC cells. ( A ) Western blot shows the knockdown of c-Myc without interfering with GLS1 expression in HepG2 and Hep3B cells. ( B ) Western blot shows that p65 knockdown reduces GLS1 protein levels in both HepG2 and Hep3B cells. ( C ) qRT-PCR indicates that p65 knockdown reduces GLS1 mRNA levels in both HepG2 and Hep3B cells. ** p

    Journal: OncoTargets and therapy

    Article Title: Nuclear factor-κB p65 regulates glutaminase 1 expression in human hepatocellular carcinoma

    doi: 10.2147/OTT.S167408

    Figure Lengend Snippet: Knockdown or suppression of p65 reduces GLS1 expression in HCC cells. ( A ) Western blot shows the knockdown of c-Myc without interfering with GLS1 expression in HepG2 and Hep3B cells. ( B ) Western blot shows that p65 knockdown reduces GLS1 protein levels in both HepG2 and Hep3B cells. ( C ) qRT-PCR indicates that p65 knockdown reduces GLS1 mRNA levels in both HepG2 and Hep3B cells. ** p

    Article Snippet: Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) Upon relative treatment, cells were lysed, and total RNA was isolated using TRIzol (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Expressing, Western Blot, Quantitative RT-PCR

    LINC01296 knockdown suppressed the migration of bladder cancer cells. Notes: ( A , B ) Transwell assays were used to measure the migration ability of T24 and 5637 cells transfected with si-LINC01296 or si-NC. ( C , D ) Wound healing assays were used to measure the migration ability of T24 and 5637 cells transfected with si-LINC01296 or si-NC. ( E ) qRT- PCR analyses were used to measure mRNA levels of E-cadherin and N-cadherin following the treatment of bladder cells with anti-LINC01296 siRNA. ( F ) Western blot assays were used to measure protein levels of E-cadherin and N-cadherin following the treatment of bladder cells with anti-LINC01296 siRNA. Data are presented as mean ± SD of three independent experiments. *P

    Journal: OncoTargets and therapy

    Article Title: Long noncoding RNA LINC01296 promotes cancer-cell proliferation and metastasis in urothelial carcinoma of the bladder

    doi: 10.2147/OTT.S192809

    Figure Lengend Snippet: LINC01296 knockdown suppressed the migration of bladder cancer cells. Notes: ( A , B ) Transwell assays were used to measure the migration ability of T24 and 5637 cells transfected with si-LINC01296 or si-NC. ( C , D ) Wound healing assays were used to measure the migration ability of T24 and 5637 cells transfected with si-LINC01296 or si-NC. ( E ) qRT- PCR analyses were used to measure mRNA levels of E-cadherin and N-cadherin following the treatment of bladder cells with anti-LINC01296 siRNA. ( F ) Western blot assays were used to measure protein levels of E-cadherin and N-cadherin following the treatment of bladder cells with anti-LINC01296 siRNA. Data are presented as mean ± SD of three independent experiments. *P

    Article Snippet: Quantitative real-time PCR (qRT-PCR) was performed on an ABI 7500 (Thermo Fisher Scientific) instrument using a KAPA SYBR Green FAST qPCR Kit (KAPA, Wilmington, MA, US) according to the manufacturer’s instructions.

    Techniques: Migration, Transfection, Quantitative RT-PCR, Western Blot

    LINC01296 knockdown inhibited the proliferation of bladder cancer cells. Notes: ( A ) Relative LINC01296 expressions in human bladder cancer cell lines (RT4, T24, and 5637). ( B , C ) MTT assays were performed to measure the proliferation viability of T24 and 5637 cells after transfection. ( D , E ) qRT-PCR results of LINC01296 expressions following the treatment of T24 and 5637 cells with anti-LINC01296 siRNA. ( F , G ) Colony formation assays were used to measure the number of clone formation of bladder cancer cells after transfection. ( H , I ) Flow cytometry assays were performed to analyze the cell cycle progression of T24 and 5637 cells after they were transfected with anti-LINC01296 siRNA. Data are presented as mean ± SD of three independent experiments. *P

    Journal: OncoTargets and therapy

    Article Title: Long noncoding RNA LINC01296 promotes cancer-cell proliferation and metastasis in urothelial carcinoma of the bladder

    doi: 10.2147/OTT.S192809

    Figure Lengend Snippet: LINC01296 knockdown inhibited the proliferation of bladder cancer cells. Notes: ( A ) Relative LINC01296 expressions in human bladder cancer cell lines (RT4, T24, and 5637). ( B , C ) MTT assays were performed to measure the proliferation viability of T24 and 5637 cells after transfection. ( D , E ) qRT-PCR results of LINC01296 expressions following the treatment of T24 and 5637 cells with anti-LINC01296 siRNA. ( F , G ) Colony formation assays were used to measure the number of clone formation of bladder cancer cells after transfection. ( H , I ) Flow cytometry assays were performed to analyze the cell cycle progression of T24 and 5637 cells after they were transfected with anti-LINC01296 siRNA. Data are presented as mean ± SD of three independent experiments. *P

    Article Snippet: Quantitative real-time PCR (qRT-PCR) was performed on an ABI 7500 (Thermo Fisher Scientific) instrument using a KAPA SYBR Green FAST qPCR Kit (KAPA, Wilmington, MA, US) according to the manufacturer’s instructions.

    Techniques: MTT Assay, Transfection, Quantitative RT-PCR, Flow Cytometry, Cytometry

    LINC01296 was upregulated in bladder cancer and the expression level of LINC01296 was correlated with clinicopathological features of bladder cancer patients. Notes: ( A ) Comparison of relative expressions of LINC01296 in cancer and normal tissues using the GEPIA database. ( B ) Comparison of relative expressions of LINC01296 in bladder cancer and adjacent normal tissues using the GEPIA database. ( C ) A microarray containing 37,000 lncRNA and 34,000 mRNA probes was used to screen differentially expressed lncRNAs in four pairs of bladder cancer patients. ( D ) Comparison of relative expressions of LINC01296 in 54 bladder cancer tissues and 24 normal bladder tissues using qRT-PCR. ( E ) Two groups were set up according to the mean expression of LINC01296 in tumor tissues. ( F ) The LINC01296 expression was significantly higher in patients with a higher tumor stage, lymph node metastasis and a higher pathologic grade. * P

    Journal: OncoTargets and therapy

    Article Title: Long noncoding RNA LINC01296 promotes cancer-cell proliferation and metastasis in urothelial carcinoma of the bladder

    doi: 10.2147/OTT.S192809

    Figure Lengend Snippet: LINC01296 was upregulated in bladder cancer and the expression level of LINC01296 was correlated with clinicopathological features of bladder cancer patients. Notes: ( A ) Comparison of relative expressions of LINC01296 in cancer and normal tissues using the GEPIA database. ( B ) Comparison of relative expressions of LINC01296 in bladder cancer and adjacent normal tissues using the GEPIA database. ( C ) A microarray containing 37,000 lncRNA and 34,000 mRNA probes was used to screen differentially expressed lncRNAs in four pairs of bladder cancer patients. ( D ) Comparison of relative expressions of LINC01296 in 54 bladder cancer tissues and 24 normal bladder tissues using qRT-PCR. ( E ) Two groups were set up according to the mean expression of LINC01296 in tumor tissues. ( F ) The LINC01296 expression was significantly higher in patients with a higher tumor stage, lymph node metastasis and a higher pathologic grade. * P

    Article Snippet: Quantitative real-time PCR (qRT-PCR) was performed on an ABI 7500 (Thermo Fisher Scientific) instrument using a KAPA SYBR Green FAST qPCR Kit (KAPA, Wilmington, MA, US) according to the manufacturer’s instructions.

    Techniques: Expressing, Microarray, Quantitative RT-PCR

    Up-regulation of miR-96 and down-regulation of FOXO1 in tumor tissues and HepG2 cells. a miR-96 relative expression in tumor tissues and non-tumor tissues by qRT-PCR; b FOXO1 mRNA relative expression in tumor tissues and non-tumor tissues by qRT-PCR; c FOXO1 protein relative expression in tumor tissues and non-tumor tissues by Western blot; d miR-96 relative expression in L02 and HepG2 cells by qRT-PCR; e FOXO1 mRNA relative expression in L02 and HepG2 cells by qRT-PCR; f FOXO1 protein relative expression in L02 and HepG2 cells by Western blot. ** P

    Journal: Cancer Cell International

    Article Title: miR-96 exerts carcinogenic effect by activating AKT/GSK-3β/β-catenin signaling pathway through targeting inhibition of FOXO1 in hepatocellular carcinoma

    doi: 10.1186/s12935-019-0756-7

    Figure Lengend Snippet: Up-regulation of miR-96 and down-regulation of FOXO1 in tumor tissues and HepG2 cells. a miR-96 relative expression in tumor tissues and non-tumor tissues by qRT-PCR; b FOXO1 mRNA relative expression in tumor tissues and non-tumor tissues by qRT-PCR; c FOXO1 protein relative expression in tumor tissues and non-tumor tissues by Western blot; d miR-96 relative expression in L02 and HepG2 cells by qRT-PCR; e FOXO1 mRNA relative expression in L02 and HepG2 cells by qRT-PCR; f FOXO1 protein relative expression in L02 and HepG2 cells by Western blot. ** P

    Article Snippet: Quantitative real time polymerase chain reaction (qRT-PCR) Total RNA in tumor tissues and cells was obtained by Trizol kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the the instruction manual.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot

    FOXO1 was directly suppressed by miR-96. a Prediction of binding sites for FOXO1 and miR-96 by Target Scan; b Luciferase reporter assay; c FOXO1 mRNA expression by qRT-PCR; d FOXO1 protein expression by Western blot. ## P

    Journal: Cancer Cell International

    Article Title: miR-96 exerts carcinogenic effect by activating AKT/GSK-3β/β-catenin signaling pathway through targeting inhibition of FOXO1 in hepatocellular carcinoma

    doi: 10.1186/s12935-019-0756-7

    Figure Lengend Snippet: FOXO1 was directly suppressed by miR-96. a Prediction of binding sites for FOXO1 and miR-96 by Target Scan; b Luciferase reporter assay; c FOXO1 mRNA expression by qRT-PCR; d FOXO1 protein expression by Western blot. ## P

    Article Snippet: Quantitative real time polymerase chain reaction (qRT-PCR) Total RNA in tumor tissues and cells was obtained by Trizol kit (Thermo Fisher Scientific, Waltham, MA, USA) according to the the instruction manual.

    Techniques: Binding Assay, Luciferase, Reporter Assay, Expressing, Quantitative RT-PCR, Western Blot

    Serum AAT c-myc Expression, Muscle Immunohistochemistry for AAT, and RT-PCR (A) Western blot gel image for all time points (please note that several AAV8 day-60 samples were missing). (B) Semi-quantitation of western blot (WB) data represented as mean ± SEM. (C) Immunohistochemistry for AAT at day-60 representative sections for each dosing route for both AAV1 and AAV8. Muscle samples were collected for immunohistochemistry (IHC) from the level of the IM injections in both the quadriceps and gastrocnemius. (D) RT-PCR data from day-60 muscle and liver tissue. The gastrocnemius muscle samples were used for all groups. IM muscle samples were collected from the injection site. IM, intramuscular; IAPD, intra-arterial push and dwell; VLP, venous limb perfusion.

    Journal: Molecular Therapy. Methods & Clinical Development

    Article Title: Bridging from Intramuscular to Limb Perfusion Delivery of rAAV: Optimization in a Non-human Primate Study

    doi: 10.1016/j.omtm.2019.01.013

    Figure Lengend Snippet: Serum AAT c-myc Expression, Muscle Immunohistochemistry for AAT, and RT-PCR (A) Western blot gel image for all time points (please note that several AAV8 day-60 samples were missing). (B) Semi-quantitation of western blot (WB) data represented as mean ± SEM. (C) Immunohistochemistry for AAT at day-60 representative sections for each dosing route for both AAV1 and AAV8. Muscle samples were collected for immunohistochemistry (IHC) from the level of the IM injections in both the quadriceps and gastrocnemius. (D) RT-PCR data from day-60 muscle and liver tissue. The gastrocnemius muscle samples were used for all groups. IM muscle samples were collected from the injection site. IM, intramuscular; IAPD, intra-arterial push and dwell; VLP, venous limb perfusion.

    Article Snippet: Real-Time qRT-PCR Frozen day-60 liver and gastrocnemius muscle samples (from dosed limbs) were used to extract RNA using TRIzol Reagent according to the manufacturer’s instructions (Thermo Fisher Scientific, #15596026).

    Techniques: Expressing, Immunohistochemistry, Reverse Transcription Polymerase Chain Reaction, Western Blot, Quantitation Assay, Injection