anti-gapdh Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Millipore anti gapdh antibody
    ( A ) Mean (±SEM) percent freezing during test for contextual fear in Experiment 1. Rats were in one of four groups: High fear ( n = 5), Low fear ( n = 6), imm Shock ( n = 8), expose only ( n = 8). ( B ) Mean (±SEM) <t>FGF2/GAPDH</t> immunoreactivity in Experiment 1. Data are presented as a percentage of mean data from group expose only. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 1. ( D ) Representative blots for each group.
    Anti Gapdh Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 6474 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh antibody/product/Millipore
    Average 99 stars, based on 6474 article reviews
    Price from $9.99 to $1999.99
    anti gapdh antibody - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    95
    BioLegend anti gapdh primary ab
    ( A ) Mean (±SEM) percent freezing during test for contextual fear in Experiment 1. Rats were in one of four groups: High fear ( n = 5), Low fear ( n = 6), imm Shock ( n = 8), expose only ( n = 8). ( B ) Mean (±SEM) <t>FGF2/GAPDH</t> immunoreactivity in Experiment 1. Data are presented as a percentage of mean data from group expose only. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 1. ( D ) Representative blots for each group.
    Anti Gapdh Primary Ab, supplied by BioLegend, used in various techniques. Bioz Stars score: 95/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh primary ab/product/BioLegend
    Average 95 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    anti gapdh primary ab - by Bioz Stars, 2020-08
    95/100 stars
      Buy from Supplier

    93
    Santa Cruz Biotechnology anti gapdh antibody
    <t>TRF2</t> binding on hTERT promoter is independent of telomere looping A . Schematic for insertion of Gaussia luciferase gene, driven by (+33 to-1276) bp hTERT promoter inserted at CCR5 safe harbour locus (46Mb away from nearest telomere using CRISPR/Cas9) in HEK293T cells. B . Effect of TRF2 silencing or over-expression of DNA binding mutants of TRF2, TRF2-DelB, TRF2-DelM and TRF2-DelB DelM on Gaussia luciferase activity, normalized over total protein C . qRT-PCR following TRF2 ChIP on the exogenously inserted hTERT promoter at CCR5 locus ; normalized over mock (IgG) ( <t>GAPDH</t> promoter-negative control for TRF2 occupancy) D . qRT-PCR following ChIP for shelterin complex proteins TRF1, POT1 and RAP1, on hTERT promoter insert in HEK293T cells E . qRT-PCR following ChIP for TRF1, POT1 and RAP1 on endogenous hTERT promoter in HT1080 cells (in D-E: Chromosome 5p (interstitial telomeric sequence) ITS site-positive control and GAPDH promoter-negative control). All error bars represent ± standard deviations from mean values and p values have been calculated by paired /un paired T-test. (*: p
    Anti Gapdh Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 2476 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh antibody/product/Santa Cruz Biotechnology
    Average 93 stars, based on 2476 article reviews
    Price from $9.99 to $1999.99
    anti gapdh antibody - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    93
    American Research Products anti gapdh antibody
    Levels of <t>ABCC4</t> in cells established using the Flp-In™ system. ABCC4 and <t>GAPDH</t> levels were determined by western blotting analysis with specific antibodies for ABCC4 and GAPDH and their levels were quantified using ImageJ (Wayne Rasband, Bethesda, MD, USA) as described in Materials and Methods. ABCC4-specific monoclonal antibody (M4I-10) or GAPDH-specific antibody was used for protein detection in PNGase F-treated cell lysate. The experiments were performed independently more than two times. Data are expressed as mean values ± S.D. (n = 3). Statistical analyses were performed using one-way ANOVA and Tukey’s HSD test (* p
    Anti Gapdh Antibody, supplied by American Research Products, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh antibody/product/American Research Products
    Average 93 stars, based on 19 article reviews
    Price from $9.99 to $1999.99
    anti gapdh antibody - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    92
    Proteintech anti gapdh antibodies
    Effects of PCNP on the apoptosis of human neuroblastoma cells. a The apoptotic levels of SH-SY5Y and SK-N-SH cells were measured by TUNEL staining; original magnification 100×. b The percentages of TUNEL-positive cells were calculated by the formula shown above. c Western blotting analysis for the expression of cleaved <t>caspase-3,</t> − 8, and − 9 and cleaved PARP in SH-SY5Y and SK-N-SH cells. <t>GAPDH</t> was used as the loading control. ( d, e ) The densitometry analysis of each factor was performed in SH-SY5Y and SK-N-SH cells, normalized to the corresponding GAPDH level. Data are presented as mean ± SEM of three independent experiments; * P
    Anti Gapdh Antibodies, supplied by Proteintech, used in various techniques. Bioz Stars score: 92/100, based on 92 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh antibodies/product/Proteintech
    Average 92 stars, based on 92 article reviews
    Price from $9.99 to $1999.99
    anti gapdh antibodies - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    92
    Becton Dickinson anti gapdh antibody
    BACE1 overexpression reduces the surface levels of Na v ). Four matching slice pairs from similar locations of the brain were selected from BACE1-transgenic (BACE1-tg) and age-matched wild-type control mice (WT), respectively. Total lysate (1/20th of total input lysate, left panel ) and NeutrAvidin captured surface biotinylated proteins ( right panel ) were analyzed by Western blot (WB) using antibodies against Na v 1 <t>α-subunits</t> (pan) and control <t>GAPDH.</t>
    Anti Gapdh Antibody, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh antibody/product/Becton Dickinson
    Average 92 stars, based on 33 article reviews
    Price from $9.99 to $1999.99
    anti gapdh antibody - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc anti gapdh antibody
    Western blots showing the effect of manganese, glutamate or riluzole or combination of the three on <t>JNK</t> phosphorylation in neuronally differentiated P19 cells. (A) Western blots showing the levels of phosphorylated JNK in differentiated P19 cells treated with the Mn (0.3 mM), glutamate (G; 5 mM) and /or riluzole (R; 10 μM) for 18 hrs.; <t>GAPDH</t> antibody was used as a loading control. (B) Graphical representation of band densities from four separate experiments. Results are presented as mean ± SE; Mn vs. control a - p
    Anti Gapdh Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1497 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh antibody/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1497 article reviews
    Price from $9.99 to $1999.99
    anti gapdh antibody - by Bioz Stars, 2020-08
    94/100 stars
      Buy from Supplier

    91
    Trevigen anti gapdh ab
    Western blots showing the effect of manganese, glutamate or riluzole or combination of the three on <t>JNK</t> phosphorylation in neuronally differentiated P19 cells. (A) Western blots showing the levels of phosphorylated JNK in differentiated P19 cells treated with the Mn (0.3 mM), glutamate (G; 5 mM) and /or riluzole (R; 10 μM) for 18 hrs.; <t>GAPDH</t> antibody was used as a loading control. (B) Graphical representation of band densities from four separate experiments. Results are presented as mean ± SE; Mn vs. control a - p
    Anti Gapdh Ab, supplied by Trevigen, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh ab/product/Trevigen
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    anti gapdh ab - by Bioz Stars, 2020-08
    91/100 stars
      Buy from Supplier

    99
    Abcam anti gapdh antibody
    miR-130b-3p participates in the regulation of Dsg1 expression. ( a ) Primary human keratinocytes were isolated as described in Materials and Methods and plated on dishes. Before they reached confluence, cells were induced to differentiate by adding 1.8 mM CaCl 2 to the culture medium. Cells were collected at the indicated time points to perform western blot analysis. Differentiation was evaluated by western blot for involucrin, <t>K10</t> and TG1 ( a ). Dsg1 protein ( b ) upregulation during differentiation is also shown; <t>GAPDH</t> is used as loading control. The histograms show the quantitative mRNA level of involucrin, K10, TG1 and Dsg1 (fold over control; black bars) as mean±S.D. from three independent experiments. ( c ) Keratinocytes were exposed to calcium stimulation and miR-130b-3p levels were analyzed by qRT-PCR. Data are shown as mean±S.D. from three independent experiments. ( d ) Putative miR-130b-3p-binding site in the 3’-UTR region of Dsg1. ( e ) miR-130b-3p suppresses the expression of Dsg1. Keratinocytes were transfected with mimic-miR-130b-3p (miR-130b-3p) or mimic negative control (mimic-NC). The levels of Dsg1 were analyzed by immunoblot. n =3. ( f ) Knockdown of miR-130b-3p increases the expression levels of Dsg1. Keratinocytes were transfected with miR-130b-3p antagomir (anta-130b-3p) or the antagomir-negative control (anta-NC), the levels of Dsg1 were analyzed by immunoblot; n =3. ( g ) Keratinocytes were treated with miR-130b-3p or mimic-NC, and cells were exposed to 1.8 mM CaCl 2 for 72 h. The levels of Dsg1 were analyzed by immunoblot. n =3. * P
    Anti Gapdh Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 2382 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh antibody/product/Abcam
    Average 99 stars, based on 2382 article reviews
    Price from $9.99 to $1999.99
    anti gapdh antibody - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    94
    Elabscience anti gapdh ab
    miR-130b-3p participates in the regulation of Dsg1 expression. ( a ) Primary human keratinocytes were isolated as described in Materials and Methods and plated on dishes. Before they reached confluence, cells were induced to differentiate by adding 1.8 mM CaCl 2 to the culture medium. Cells were collected at the indicated time points to perform western blot analysis. Differentiation was evaluated by western blot for involucrin, <t>K10</t> and TG1 ( a ). Dsg1 protein ( b ) upregulation during differentiation is also shown; <t>GAPDH</t> is used as loading control. The histograms show the quantitative mRNA level of involucrin, K10, TG1 and Dsg1 (fold over control; black bars) as mean±S.D. from three independent experiments. ( c ) Keratinocytes were exposed to calcium stimulation and miR-130b-3p levels were analyzed by qRT-PCR. Data are shown as mean±S.D. from three independent experiments. ( d ) Putative miR-130b-3p-binding site in the 3’-UTR region of Dsg1. ( e ) miR-130b-3p suppresses the expression of Dsg1. Keratinocytes were transfected with mimic-miR-130b-3p (miR-130b-3p) or mimic negative control (mimic-NC). The levels of Dsg1 were analyzed by immunoblot. n =3. ( f ) Knockdown of miR-130b-3p increases the expression levels of Dsg1. Keratinocytes were transfected with miR-130b-3p antagomir (anta-130b-3p) or the antagomir-negative control (anta-NC), the levels of Dsg1 were analyzed by immunoblot; n =3. ( g ) Keratinocytes were treated with miR-130b-3p or mimic-NC, and cells were exposed to 1.8 mM CaCl 2 for 72 h. The levels of Dsg1 were analyzed by immunoblot. n =3. * P
    Anti Gapdh Ab, supplied by Elabscience, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh ab/product/Elabscience
    Average 94 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    anti gapdh ab - by Bioz Stars, 2020-08
    94/100 stars
      Buy from Supplier

    80
    Charles River Laboratories anti gapdh polyclonal ab
    miR-130b-3p participates in the regulation of Dsg1 expression. ( a ) Primary human keratinocytes were isolated as described in Materials and Methods and plated on dishes. Before they reached confluence, cells were induced to differentiate by adding 1.8 mM CaCl 2 to the culture medium. Cells were collected at the indicated time points to perform western blot analysis. Differentiation was evaluated by western blot for involucrin, <t>K10</t> and TG1 ( a ). Dsg1 protein ( b ) upregulation during differentiation is also shown; <t>GAPDH</t> is used as loading control. The histograms show the quantitative mRNA level of involucrin, K10, TG1 and Dsg1 (fold over control; black bars) as mean±S.D. from three independent experiments. ( c ) Keratinocytes were exposed to calcium stimulation and miR-130b-3p levels were analyzed by qRT-PCR. Data are shown as mean±S.D. from three independent experiments. ( d ) Putative miR-130b-3p-binding site in the 3’-UTR region of Dsg1. ( e ) miR-130b-3p suppresses the expression of Dsg1. Keratinocytes were transfected with mimic-miR-130b-3p (miR-130b-3p) or mimic negative control (mimic-NC). The levels of Dsg1 were analyzed by immunoblot. n =3. ( f ) Knockdown of miR-130b-3p increases the expression levels of Dsg1. Keratinocytes were transfected with miR-130b-3p antagomir (anta-130b-3p) or the antagomir-negative control (anta-NC), the levels of Dsg1 were analyzed by immunoblot; n =3. ( g ) Keratinocytes were treated with miR-130b-3p or mimic-NC, and cells were exposed to 1.8 mM CaCl 2 for 72 h. The levels of Dsg1 were analyzed by immunoblot. n =3. * P
    Anti Gapdh Polyclonal Ab, supplied by Charles River Laboratories, used in various techniques. Bioz Stars score: 80/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti gapdh polyclonal ab/product/Charles River Laboratories
    Average 80 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    anti gapdh polyclonal ab - by Bioz Stars, 2020-08
    80/100 stars
      Buy from Supplier

    Image Search Results


    ( A ) Mean (±SEM) percent freezing during test for contextual fear in Experiment 1. Rats were in one of four groups: High fear ( n = 5), Low fear ( n = 6), imm Shock ( n = 8), expose only ( n = 8). ( B ) Mean (±SEM) FGF2/GAPDH immunoreactivity in Experiment 1. Data are presented as a percentage of mean data from group expose only. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 1. ( D ) Representative blots for each group.

    Journal: Learning & Memory

    Article Title: Individual differences in the expression of conditioned fear are associated with endogenous fibroblast growth factor 2

    doi: 10.1101/lm.039644.115

    Figure Lengend Snippet: ( A ) Mean (±SEM) percent freezing during test for contextual fear in Experiment 1. Rats were in one of four groups: High fear ( n = 5), Low fear ( n = 6), imm Shock ( n = 8), expose only ( n = 8). ( B ) Mean (±SEM) FGF2/GAPDH immunoreactivity in Experiment 1. Data are presented as a percentage of mean data from group expose only. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 1. ( D ) Representative blots for each group.

    Article Snippet: Proteins were transferred to nitrocellulose membranes, and nonspecific immunoreactivity was blocked with 5% nonfat dry milk/3% BSA in TBST for 60 min. Membranes were incubated overnight at 4°C with anti-FGF2 antibody (1:500; Abcam) or anti-GAPDH antibody (1:500; Millipore).

    Techniques:

    ( A ) Mean (±SEM) percent CS-elicited freezing in Experiment 2. Rats were in one of three groups: High fear ( n = 8), Low fear ( n = 9), CS only ( n = 5). ( B ) Mean (±SEM) FGF2/GAPDH immunoreactivity in Experiment 2. Data are presented as a percentage of mean data from group CS only. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 2. ( D ) Representative blots for each group.

    Journal: Learning & Memory

    Article Title: Individual differences in the expression of conditioned fear are associated with endogenous fibroblast growth factor 2

    doi: 10.1101/lm.039644.115

    Figure Lengend Snippet: ( A ) Mean (±SEM) percent CS-elicited freezing in Experiment 2. Rats were in one of three groups: High fear ( n = 8), Low fear ( n = 9), CS only ( n = 5). ( B ) Mean (±SEM) FGF2/GAPDH immunoreactivity in Experiment 2. Data are presented as a percentage of mean data from group CS only. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 2. ( D ) Representative blots for each group.

    Article Snippet: Proteins were transferred to nitrocellulose membranes, and nonspecific immunoreactivity was blocked with 5% nonfat dry milk/3% BSA in TBST for 60 min. Membranes were incubated overnight at 4°C with anti-FGF2 antibody (1:500; Abcam) or anti-GAPDH antibody (1:500; Millipore).

    Techniques:

    ( A ) Mean (±SEM) percent freezing during test for contextual fear in Experiment 3. Rats were in one of three groups: High fear ( n = 13), Low fear ( n = 13), MK801 ( n = 12). ( B ) Mean (±SEM) FGF2/GAPDH immunoreactivity in Experiment 3. Data are presented as a percentage of mean data from group MK801. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 3. ( D ) Representative blots for each group.

    Journal: Learning & Memory

    Article Title: Individual differences in the expression of conditioned fear are associated with endogenous fibroblast growth factor 2

    doi: 10.1101/lm.039644.115

    Figure Lengend Snippet: ( A ) Mean (±SEM) percent freezing during test for contextual fear in Experiment 3. Rats were in one of three groups: High fear ( n = 13), Low fear ( n = 13), MK801 ( n = 12). ( B ) Mean (±SEM) FGF2/GAPDH immunoreactivity in Experiment 3. Data are presented as a percentage of mean data from group MK801. ( C ) Correlation between FGF2/GAPDH immunoreactivity and percent freezing for Experiment 3. ( D ) Representative blots for each group.

    Article Snippet: Proteins were transferred to nitrocellulose membranes, and nonspecific immunoreactivity was blocked with 5% nonfat dry milk/3% BSA in TBST for 60 min. Membranes were incubated overnight at 4°C with anti-FGF2 antibody (1:500; Abcam) or anti-GAPDH antibody (1:500; Millipore).

    Techniques:

    Inhibition of miR-103 upregulates TIMP-3 protein expression in endometrial cancer cells. Ishikawa and HEC-1B cells were transfected with anti-miR-103 (Anti-miR) or negative control oligonucleotide (Neg Control) and the TIMP-3 protein level was detected by western blot analysis. The GAPDH protein was regarded as an endogenous normalizer. (A and C) Inhibition of miR-103 upregulates the endogenous TIMP-3 protein expression in Ishikawa and HEC-1B cells. (B and D) A significant increase in TIMP-3 activity was detected in Ishikawa and HEC-1B cells transfected with anti-miR-103, compared with the other two groups. Each bar is the mean ± SEM from three independent experiments. * P

    Journal: Oncology Letters

    Article Title: microRNA-103 regulates the growth and invasion of endometrial cancer cells through the downregulation of tissue inhibitor of metalloproteinase 3

    doi: 10.3892/ol.2012.638

    Figure Lengend Snippet: Inhibition of miR-103 upregulates TIMP-3 protein expression in endometrial cancer cells. Ishikawa and HEC-1B cells were transfected with anti-miR-103 (Anti-miR) or negative control oligonucleotide (Neg Control) and the TIMP-3 protein level was detected by western blot analysis. The GAPDH protein was regarded as an endogenous normalizer. (A and C) Inhibition of miR-103 upregulates the endogenous TIMP-3 protein expression in Ishikawa and HEC-1B cells. (B and D) A significant increase in TIMP-3 activity was detected in Ishikawa and HEC-1B cells transfected with anti-miR-103, compared with the other two groups. Each bar is the mean ± SEM from three independent experiments. * P

    Article Snippet: The TIMP-3 antibody was purchased from Abcam (Cambridge, MA, USA), GAPDH antibody from Sigma (Steinheim, Germany) and goat anti-mouse IgG from Bio-Rad (Hercules, CA, USA).

    Techniques: Inhibition, Expressing, Transfection, Negative Control, Western Blot, Activity Assay

    In vitro human RPE GAPDH and nitrotyrosine. Cultured human RPE cells (2 nd passage) were exposed to control medium (A,C) or medium containing 200 μM hydrogen peroxide (B,D) for 24 hours. Immumocytochemistry for GAPDH (A,B) and nitrotyrosine (C,D)

    Journal: Redox report : communications in free radical research

    Article Title: Ischemia-induced nitrotyrosine formation and nuclear translocation of glyceraldehyde-3-phosphate dehydrogenase in human retinal pigment epithelium in vivo

    doi: 10.1179/174329211X12968219310710

    Figure Lengend Snippet: In vitro human RPE GAPDH and nitrotyrosine. Cultured human RPE cells (2 nd passage) were exposed to control medium (A,C) or medium containing 200 μM hydrogen peroxide (B,D) for 24 hours. Immumocytochemistry for GAPDH (A,B) and nitrotyrosine (C,D)

    Article Snippet: After blocking procedures, sections were incubated with 2 μg/mL mouse monoclonal anti-nitrotyrosine antibody (1:50; Millipore, Billerica, MA), monoclonal anti-GAPDH antibody (1:50; Millipore) or isotype control IgG diluted in 1% BSA in PBS overnight at 4°C.

    Techniques: In Vitro, Cell Culture

    In vivo human RPE and choroidal GAPDH and nitrotyrosine. Ischemic choroid and RPE (A) demonstrate GAPDH nuclear translocation in RPE and vascular endothelial cells (C) as well as considerable immunoreactive nitrotyrosine (E). RPE and choroid of control

    Journal: Redox report : communications in free radical research

    Article Title: Ischemia-induced nitrotyrosine formation and nuclear translocation of glyceraldehyde-3-phosphate dehydrogenase in human retinal pigment epithelium in vivo

    doi: 10.1179/174329211X12968219310710

    Figure Lengend Snippet: In vivo human RPE and choroidal GAPDH and nitrotyrosine. Ischemic choroid and RPE (A) demonstrate GAPDH nuclear translocation in RPE and vascular endothelial cells (C) as well as considerable immunoreactive nitrotyrosine (E). RPE and choroid of control

    Article Snippet: After blocking procedures, sections were incubated with 2 μg/mL mouse monoclonal anti-nitrotyrosine antibody (1:50; Millipore, Billerica, MA), monoclonal anti-GAPDH antibody (1:50; Millipore) or isotype control IgG diluted in 1% BSA in PBS overnight at 4°C.

    Techniques: In Vivo, Translocation Assay

    Generation and characterization of apoE-fKI/Dlx-Cre mice. A , B , Expression of ZsGreen1 (green) in the cortex and hippocampus of Dlx-Cre-positive ( A ) and -negative ( B ) ZsGreen1 reporter mice. Scale bars: 250 μm. C–F , ZsGreen1 was expressed in GABA-positive ( C , E ) and somatostatin (SOM)-positive ( D , F ) inhibitory interneurons in the cortex and hippocampus of Dlx-Cre-positive apoE-fKI mice. Scale bars: 15 μm. G , H , Anti-apoE immunostaining revealed the presence of apoE protein in GABA-positive hilar interneurons in aged apoE4-fKI mice ( G ), but not in apoE4-fKI/Dlx-Cre mice ( H ). Scale bars: 15 μm. I , Images of GABA-positive hilar interneurons before and after laser capture. Inhibitory interneuron were identified by anti-GABA immunostaining. Arrows indicate the cell before and after laser capture. Scale bars: 15 μm. J , Schematic of primers and their binding sites on the human APOE gene. K , PCR with primers 1 and 2 resulted in an amplified product in samples of apoE4-fKI mice, but not in samples of apoE4-fKI/Dlx-Cre mice (left). PCR with primers 1 and 3 resulted in an amplified product in samples of apoE4-fKI/Dlx-Cre mice, but not in samples of apoE4-fKI mice (right). Fifty nuclei per sample were used. L , Representative fluorescent Western blot of apoE (green) and GAPDH (red) in cortical and hippocampal lysates of 17-month-old female mice with different genotypes. M , N , Quantification of apoE protein levels relative to GAPDH protein levels in cortical ( M ) and hippocampal lysates ( N ) of 17-month-old mice ( n = 5 per genotype). For both the cortex and the hippocampus, the apoE level in apoE3-fKI mice was normalized to 1, and apoE levels in other groups of mice were presented relative to those in apoE3-fKI mice. * p

    Journal: The Journal of Neuroscience

    Article Title: Apolipoprotein E4 Produced in GABAergic Interneurons Causes Learning and Memory Deficits in Mice

    doi: 10.1523/JNEUROSCI.2281-14.2014

    Figure Lengend Snippet: Generation and characterization of apoE-fKI/Dlx-Cre mice. A , B , Expression of ZsGreen1 (green) in the cortex and hippocampus of Dlx-Cre-positive ( A ) and -negative ( B ) ZsGreen1 reporter mice. Scale bars: 250 μm. C–F , ZsGreen1 was expressed in GABA-positive ( C , E ) and somatostatin (SOM)-positive ( D , F ) inhibitory interneurons in the cortex and hippocampus of Dlx-Cre-positive apoE-fKI mice. Scale bars: 15 μm. G , H , Anti-apoE immunostaining revealed the presence of apoE protein in GABA-positive hilar interneurons in aged apoE4-fKI mice ( G ), but not in apoE4-fKI/Dlx-Cre mice ( H ). Scale bars: 15 μm. I , Images of GABA-positive hilar interneurons before and after laser capture. Inhibitory interneuron were identified by anti-GABA immunostaining. Arrows indicate the cell before and after laser capture. Scale bars: 15 μm. J , Schematic of primers and their binding sites on the human APOE gene. K , PCR with primers 1 and 2 resulted in an amplified product in samples of apoE4-fKI mice, but not in samples of apoE4-fKI/Dlx-Cre mice (left). PCR with primers 1 and 3 resulted in an amplified product in samples of apoE4-fKI/Dlx-Cre mice, but not in samples of apoE4-fKI mice (right). Fifty nuclei per sample were used. L , Representative fluorescent Western blot of apoE (green) and GAPDH (red) in cortical and hippocampal lysates of 17-month-old female mice with different genotypes. M , N , Quantification of apoE protein levels relative to GAPDH protein levels in cortical ( M ) and hippocampal lysates ( N ) of 17-month-old mice ( n = 5 per genotype). For both the cortex and the hippocampus, the apoE level in apoE3-fKI mice was normalized to 1, and apoE levels in other groups of mice were presented relative to those in apoE3-fKI mice. * p

    Article Snippet: Nonspecific binding sites were blocked with blocking buffer (LI-COR Biosciences) for 1 h. Membranes were incubated with polyclonal goat anti-apoE antibody (1:5000; Calbiochem) and monoclonal mouse anti-GAPDH antibody (1:10,000; Millipore Bioscience Research Reagents) overnight at 4°C.

    Techniques: Mouse Assay, Expressing, Immunostaining, Binding Assay, Polymerase Chain Reaction, Amplification, Western Blot

    Generation and characterization of apoE-fKI/Syn-1-Cre mice. A , B , Expression of ZsGreen1 (green) in the cortex and hippocampus of Syn-1-Cre-positive ( A ) and -negative ( B ) ZsGreen1 reporter mice. Scale bars: 250 μm. C–H , ZsGreen1 was expressed in NeuN-positive neurons ( C , F ) and GABA-positive inhibitory interneurons ( E , H ), but not in GFAP-positive astrocytes ( D , G ) in the cortex and hippocampus of Syn-1-Cre-positive apoE-fKI mice. Scale bars: 30 μm. I , Images of the dentate granular cell layer before and after laser capture. Neuronal nuclei were identified by anti-NeuN immunostaining. Arrows indicate the cell before and after laser capture. Scale bars: 30 μm. J , Schematic of primers and their binding sites on the human APOE gene. K , PCR with primers 1 and 2 resulted in an amplified product in samples of apoE4-fKI mice, but not in samples of apoE4-fKI/Syn-1-Cre mice (left). PCR with primers 1 and 3 resulted in an amplified product in samples of apoE4-fKI/Syn-1-Cre mice, but not in samples of apoE4-fKI mice (right). Fifty nuclei per sample were used. L , Representative fluorescent Western blot of apoE (green) and GAPDH (red) in cortical and hippocampal lysates of 17-month-old female mice with different genotypes. M , N , Quantification of apoE protein levels relative to GAPDH protein levels in cortical ( M ) and hippocampal lysates ( N ) of 17-month-old mice ( n = 5 per genotype). ApoE levels in apoE3-fKI mice were normalized to 1, and apoE levels in other groups of mice were presented relative to those in apoE3-fKI mice. * p

    Journal: The Journal of Neuroscience

    Article Title: Apolipoprotein E4 Produced in GABAergic Interneurons Causes Learning and Memory Deficits in Mice

    doi: 10.1523/JNEUROSCI.2281-14.2014

    Figure Lengend Snippet: Generation and characterization of apoE-fKI/Syn-1-Cre mice. A , B , Expression of ZsGreen1 (green) in the cortex and hippocampus of Syn-1-Cre-positive ( A ) and -negative ( B ) ZsGreen1 reporter mice. Scale bars: 250 μm. C–H , ZsGreen1 was expressed in NeuN-positive neurons ( C , F ) and GABA-positive inhibitory interneurons ( E , H ), but not in GFAP-positive astrocytes ( D , G ) in the cortex and hippocampus of Syn-1-Cre-positive apoE-fKI mice. Scale bars: 30 μm. I , Images of the dentate granular cell layer before and after laser capture. Neuronal nuclei were identified by anti-NeuN immunostaining. Arrows indicate the cell before and after laser capture. Scale bars: 30 μm. J , Schematic of primers and their binding sites on the human APOE gene. K , PCR with primers 1 and 2 resulted in an amplified product in samples of apoE4-fKI mice, but not in samples of apoE4-fKI/Syn-1-Cre mice (left). PCR with primers 1 and 3 resulted in an amplified product in samples of apoE4-fKI/Syn-1-Cre mice, but not in samples of apoE4-fKI mice (right). Fifty nuclei per sample were used. L , Representative fluorescent Western blot of apoE (green) and GAPDH (red) in cortical and hippocampal lysates of 17-month-old female mice with different genotypes. M , N , Quantification of apoE protein levels relative to GAPDH protein levels in cortical ( M ) and hippocampal lysates ( N ) of 17-month-old mice ( n = 5 per genotype). ApoE levels in apoE3-fKI mice were normalized to 1, and apoE levels in other groups of mice were presented relative to those in apoE3-fKI mice. * p

    Article Snippet: Nonspecific binding sites were blocked with blocking buffer (LI-COR Biosciences) for 1 h. Membranes were incubated with polyclonal goat anti-apoE antibody (1:5000; Calbiochem) and monoclonal mouse anti-GAPDH antibody (1:10,000; Millipore Bioscience Research Reagents) overnight at 4°C.

    Techniques: Mouse Assay, Expressing, Immunostaining, Binding Assay, Polymerase Chain Reaction, Amplification, Western Blot

    Generation and characterization of apoE-fKI/GFAP-Cre mice. A , B , Representative images of fluorescent immunostaining with anti-Cre recombinase (green) and anti-GFAP (red) in the cortex ( A ) and hippocampus ( B ) of apoE-fKI/GFAP-Cre mice. C , D , Neurons immunostained with anti-NeuN (red) did not express Cre-recombinase in the cortex ( C ) or hippocampus ( D ) of apoE-fKI/GFAP-Cre mice. E , F , Anti-apoE (green) and anti-GFAP (red) double immunostaining revealed that apoE expression was dramatically decreased in cortical ( E ) and hippocampal ( F ) astrocytes in apoE3-fKI/GFAP-Cre and apoE4-fKI/GFAP-Cre mice. G , Representative fluorescent Western blot of apoE (green) and GAPDH (red) in cortical and hippocampal lysates of 17-month-old female mice with different genotypes. H , I , Quantification of apoE protein levels relative to GAPDH in cortical ( H ) and hippocampal lysates ( I ) of 17-month-old mice ( n = 5/genotype). ApoE levels in apoE3-fKI mice were normalized to 1, and apoE levels in other groups of mice were presented relative to those in apoE3-fKI mice. *** p

    Journal: The Journal of Neuroscience

    Article Title: Apolipoprotein E4 Produced in GABAergic Interneurons Causes Learning and Memory Deficits in Mice

    doi: 10.1523/JNEUROSCI.2281-14.2014

    Figure Lengend Snippet: Generation and characterization of apoE-fKI/GFAP-Cre mice. A , B , Representative images of fluorescent immunostaining with anti-Cre recombinase (green) and anti-GFAP (red) in the cortex ( A ) and hippocampus ( B ) of apoE-fKI/GFAP-Cre mice. C , D , Neurons immunostained with anti-NeuN (red) did not express Cre-recombinase in the cortex ( C ) or hippocampus ( D ) of apoE-fKI/GFAP-Cre mice. E , F , Anti-apoE (green) and anti-GFAP (red) double immunostaining revealed that apoE expression was dramatically decreased in cortical ( E ) and hippocampal ( F ) astrocytes in apoE3-fKI/GFAP-Cre and apoE4-fKI/GFAP-Cre mice. G , Representative fluorescent Western blot of apoE (green) and GAPDH (red) in cortical and hippocampal lysates of 17-month-old female mice with different genotypes. H , I , Quantification of apoE protein levels relative to GAPDH in cortical ( H ) and hippocampal lysates ( I ) of 17-month-old mice ( n = 5/genotype). ApoE levels in apoE3-fKI mice were normalized to 1, and apoE levels in other groups of mice were presented relative to those in apoE3-fKI mice. *** p

    Article Snippet: Nonspecific binding sites were blocked with blocking buffer (LI-COR Biosciences) for 1 h. Membranes were incubated with polyclonal goat anti-apoE antibody (1:5000; Calbiochem) and monoclonal mouse anti-GAPDH antibody (1:10,000; Millipore Bioscience Research Reagents) overnight at 4°C.

    Techniques: Mouse Assay, Immunostaining, Double Immunostaining, Expressing, Western Blot

    TRF2 binding on hTERT promoter is independent of telomere looping A . Schematic for insertion of Gaussia luciferase gene, driven by (+33 to-1276) bp hTERT promoter inserted at CCR5 safe harbour locus (46Mb away from nearest telomere using CRISPR/Cas9) in HEK293T cells. B . Effect of TRF2 silencing or over-expression of DNA binding mutants of TRF2, TRF2-DelB, TRF2-DelM and TRF2-DelB DelM on Gaussia luciferase activity, normalized over total protein C . qRT-PCR following TRF2 ChIP on the exogenously inserted hTERT promoter at CCR5 locus ; normalized over mock (IgG) ( GAPDH promoter-negative control for TRF2 occupancy) D . qRT-PCR following ChIP for shelterin complex proteins TRF1, POT1 and RAP1, on hTERT promoter insert in HEK293T cells E . qRT-PCR following ChIP for TRF1, POT1 and RAP1 on endogenous hTERT promoter in HT1080 cells (in D-E: Chromosome 5p (interstitial telomeric sequence) ITS site-positive control and GAPDH promoter-negative control). All error bars represent ± standard deviations from mean values and p values have been calculated by paired /un paired T-test. (*: p

    Journal: bioRxiv

    Article Title: Human Telomerase Expression is under Direct Transcriptional Control of the Telomere-binding-factor TRF2

    doi: 10.1101/2020.01.15.907626

    Figure Lengend Snippet: TRF2 binding on hTERT promoter is independent of telomere looping A . Schematic for insertion of Gaussia luciferase gene, driven by (+33 to-1276) bp hTERT promoter inserted at CCR5 safe harbour locus (46Mb away from nearest telomere using CRISPR/Cas9) in HEK293T cells. B . Effect of TRF2 silencing or over-expression of DNA binding mutants of TRF2, TRF2-DelB, TRF2-DelM and TRF2-DelB DelM on Gaussia luciferase activity, normalized over total protein C . qRT-PCR following TRF2 ChIP on the exogenously inserted hTERT promoter at CCR5 locus ; normalized over mock (IgG) ( GAPDH promoter-negative control for TRF2 occupancy) D . qRT-PCR following ChIP for shelterin complex proteins TRF1, POT1 and RAP1, on hTERT promoter insert in HEK293T cells E . qRT-PCR following ChIP for TRF1, POT1 and RAP1 on endogenous hTERT promoter in HT1080 cells (in D-E: Chromosome 5p (interstitial telomeric sequence) ITS site-positive control and GAPDH promoter-negative control). All error bars represent ± standard deviations from mean values and p values have been calculated by paired /un paired T-test. (*: p

    Article Snippet: After blocking the membrane was incubated with primary antibodies-anti-TRF2 antibody (Novus Biological), anti-TERT antibody (Abcam), anti-REST(Millipore), anti-EZH2(CST) and anti-GAPDH antibody (Santa-cruz).

    Techniques: Binding Assay, Luciferase, CRISPR, Over Expression, Activity Assay, Quantitative RT-PCR, Chromatin Immunoprecipitation, Negative Control, Sequencing, Positive Control

    Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Slit2-Robo4 pathway modulates LPS-induced endothelial inflammation and its expression is dysregulated during endotoxemia

    doi: 10.4049/jimmunol.1302021

    Figure Lengend Snippet: Slit2-N inhibited the LPS-induced cytokine and cell adhesion molecule ICAM-1 expression in HUVECs as well as monocyte adhesion on HUVECs HUVECs were treated with 100ng/mL LPS for 2 h (A) or 12 h (B) with or without treatment of Slit2-N. (A) Cells were stimulated by LPS with pre-treatment of a gradient of 3 nmol/L, 30 nmol/L and 60 nmol/L Slit2-N or post-treatment with 30 nmol/L Slit2-N (post-Slit2-N). Total RNA was isolated and qRT-PCR was performed to detect mRNA expression level of MCP-1 and GM-CSF. Values are normalized to control group, and are not comparable between cytokines. (B) Cell culture supernatants were collected after Slit2-N (30 nmol/L) plus LPS treatment. Cytokines secreted in the supernatants were detected with R D cytokine array kit. Values are net optical signaling intensity. (C) HUVECs were treated with 100 ng/mL LPS for 12 h with or without pre-treatment of Slit2-N (30 nmol/L). Cells were lysed and WB was performed using ICAM-1, VCAM-1 and GAPDH antibodies. (D) HUVECs were stimulated with LPS (100 ng/mL) for 6 h with or without Slit2-N (30 nmol/L). CFSE labeled THP-1 monocytic cells were added onto treated HUVEC. And after washing, attached THP-1 cells were quantified by mean fluorescence intensity at 528nm. (E) HMVECs were stimulated with 100ng/mL LPS in the presence or absence of 30 nmol/L Slit2-N for 12 h. Supernatant MCP-1 secretion was analyzed by ELISA. (F) ICAM-1 expression was detected by qRT-PCR in HMVECs with LPS stimulation in the presence or absence of Slit2-N. (* p

    Article Snippet: ICAM-1, VCAM-1 and GAPDH antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA).

    Techniques: Expressing, Isolation, Quantitative RT-PCR, Cell Culture, Western Blot, Labeling, Fluorescence, Enzyme-linked Immunosorbent Assay

    Levels of ABCC4 in cells established using the Flp-In™ system. ABCC4 and GAPDH levels were determined by western blotting analysis with specific antibodies for ABCC4 and GAPDH and their levels were quantified using ImageJ (Wayne Rasband, Bethesda, MD, USA) as described in Materials and Methods. ABCC4-specific monoclonal antibody (M4I-10) or GAPDH-specific antibody was used for protein detection in PNGase F-treated cell lysate. The experiments were performed independently more than two times. Data are expressed as mean values ± S.D. (n = 3). Statistical analyses were performed using one-way ANOVA and Tukey’s HSD test (* p

    Journal: Cells

    Article Title: A Human ABC Transporter ABCC4 Gene SNP (rs11568658, 559 G > T, G187W) Reduces ABCC4-Dependent Drug Resistance

    doi: 10.3390/cells8010039

    Figure Lengend Snippet: Levels of ABCC4 in cells established using the Flp-In™ system. ABCC4 and GAPDH levels were determined by western blotting analysis with specific antibodies for ABCC4 and GAPDH and their levels were quantified using ImageJ (Wayne Rasband, Bethesda, MD, USA) as described in Materials and Methods. ABCC4-specific monoclonal antibody (M4I-10) or GAPDH-specific antibody was used for protein detection in PNGase F-treated cell lysate. The experiments were performed independently more than two times. Data are expressed as mean values ± S.D. (n = 3). Statistical analyses were performed using one-way ANOVA and Tukey’s HSD test (* p

    Article Snippet: The membranes were incubated with monoclonal anti-ABCC4 antibody (M4I-10; GeneTex Inc., Alton Parkway Irvine, CA, USA) or anti-GAPDH antibody (anti-GAPDH-Clone 6C5 mouse monoclonal, igG2b; American Research Products, Inc., Waltham, MA, USA) at 1:1000 dilution in TBST containing 5% (w /v ) skim milk powder for 1 h at room temperature with shaking after rinsing with TBST.

    Techniques: Western Blot

    Levels of ABCC4 mRNA in cells established using the Flp-In™ system. The levels of ABCC4 and GAPDH mRNA were measured using qPCR with specific primer sets for ABCC4 and GAPDH , as described in Materials and Methods. Data are calculated as ratios relative to the GAPDH mRNA levels in the cells and normalized to the ratio of ABCC4 / GAPDH . Data are expressed as mean values ± S.D. (n = 5). Statistical analyses were performed using one-way ANOVA and Tukey’s HSD test (* p

    Journal: Cells

    Article Title: A Human ABC Transporter ABCC4 Gene SNP (rs11568658, 559 G > T, G187W) Reduces ABCC4-Dependent Drug Resistance

    doi: 10.3390/cells8010039

    Figure Lengend Snippet: Levels of ABCC4 mRNA in cells established using the Flp-In™ system. The levels of ABCC4 and GAPDH mRNA were measured using qPCR with specific primer sets for ABCC4 and GAPDH , as described in Materials and Methods. Data are calculated as ratios relative to the GAPDH mRNA levels in the cells and normalized to the ratio of ABCC4 / GAPDH . Data are expressed as mean values ± S.D. (n = 5). Statistical analyses were performed using one-way ANOVA and Tukey’s HSD test (* p

    Article Snippet: The membranes were incubated with monoclonal anti-ABCC4 antibody (M4I-10; GeneTex Inc., Alton Parkway Irvine, CA, USA) or anti-GAPDH antibody (anti-GAPDH-Clone 6C5 mouse monoclonal, igG2b; American Research Products, Inc., Waltham, MA, USA) at 1:1000 dilution in TBST containing 5% (w /v ) skim milk powder for 1 h at room temperature with shaking after rinsing with TBST.

    Techniques: Real-time Polymerase Chain Reaction

    Effects of PCNP on the apoptosis of human neuroblastoma cells. a The apoptotic levels of SH-SY5Y and SK-N-SH cells were measured by TUNEL staining; original magnification 100×. b The percentages of TUNEL-positive cells were calculated by the formula shown above. c Western blotting analysis for the expression of cleaved caspase-3, − 8, and − 9 and cleaved PARP in SH-SY5Y and SK-N-SH cells. GAPDH was used as the loading control. ( d, e ) The densitometry analysis of each factor was performed in SH-SY5Y and SK-N-SH cells, normalized to the corresponding GAPDH level. Data are presented as mean ± SEM of three independent experiments; * P

    Journal: BMC Cancer

    Article Title: PEST-containing nuclear protein mediates the proliferation, migration, and invasion of human neuroblastoma cells through MAPK and PI3K/AKT/mTOR signaling pathways

    doi: 10.1186/s12885-018-4391-9

    Figure Lengend Snippet: Effects of PCNP on the apoptosis of human neuroblastoma cells. a The apoptotic levels of SH-SY5Y and SK-N-SH cells were measured by TUNEL staining; original magnification 100×. b The percentages of TUNEL-positive cells were calculated by the formula shown above. c Western blotting analysis for the expression of cleaved caspase-3, − 8, and − 9 and cleaved PARP in SH-SY5Y and SK-N-SH cells. GAPDH was used as the loading control. ( d, e ) The densitometry analysis of each factor was performed in SH-SY5Y and SK-N-SH cells, normalized to the corresponding GAPDH level. Data are presented as mean ± SEM of three independent experiments; * P

    Article Snippet: Anti-PCNP, Anti-B-cell lymphoma-2 (Bcl-2), anti-Bcl-2-associated X protein (Bax), anti-B-cell lymphoma-extra large (Bcl-xl), anti-Bcl-xl/Bcl-2-associated death promoter (Bad), anti-cleaved caspase-3, anti-cleaved caspase-8, anti-cleaved caspase-9, anti-Cleaved poly adenosine diphosphate-ribose polymerase (PARP), and anti-GAPDH antibodies were purchased from ProteinTech (Chicago, IL, USA).

    Techniques: TUNEL Assay, Staining, Western Blot, Expressing

    BACE1 overexpression reduces the surface levels of Na v ). Four matching slice pairs from similar locations of the brain were selected from BACE1-transgenic (BACE1-tg) and age-matched wild-type control mice (WT), respectively. Total lysate (1/20th of total input lysate, left panel ) and NeutrAvidin captured surface biotinylated proteins ( right panel ) were analyzed by Western blot (WB) using antibodies against Na v 1 α-subunits (pan) and control GAPDH.

    Journal: Methods in Molecular Biology (Clifton, N.j.)

    Article Title: Surface Trafficking of Sodium Channels in Cells and in Hippocampal Slices

    doi: 10.1007/978-1-61779-328-8_23

    Figure Lengend Snippet: BACE1 overexpression reduces the surface levels of Na v ). Four matching slice pairs from similar locations of the brain were selected from BACE1-transgenic (BACE1-tg) and age-matched wild-type control mice (WT), respectively. Total lysate (1/20th of total input lysate, left panel ) and NeutrAvidin captured surface biotinylated proteins ( right panel ) were analyzed by Western blot (WB) using antibodies against Na v 1 α-subunits (pan) and control GAPDH.

    Article Snippet: Primary antibody solutions used in our study: Anti-pan α-subunit antibody (1:500, Sigma), anti-Nav 1.1 α-subunit antibody (1:500, Sigma), anti-Nav 1.2 α-subunit antibody (1:500, Alomone, Isael), anti-GAPDH antibody (BD bioscience), anti- N -cadherin antibody (BD bioscience/transduction laboratory).

    Techniques: Over Expression, Transgenic Assay, Mouse Assay, Western Blot

    Western blots showing the effect of manganese, glutamate or riluzole or combination of the three on JNK phosphorylation in neuronally differentiated P19 cells. (A) Western blots showing the levels of phosphorylated JNK in differentiated P19 cells treated with the Mn (0.3 mM), glutamate (G; 5 mM) and /or riluzole (R; 10 μM) for 18 hrs.; GAPDH antibody was used as a loading control. (B) Graphical representation of band densities from four separate experiments. Results are presented as mean ± SE; Mn vs. control a - p

    Journal: Neurochemistry International

    Article Title: Effect of Glutamate and Riluzole on Manganese-Induced Apoptotic Cell Signaling in Neuronally Differentiated Mouse P19 Cells

    doi: 10.1016/j.neuint.2012.04.015

    Figure Lengend Snippet: Western blots showing the effect of manganese, glutamate or riluzole or combination of the three on JNK phosphorylation in neuronally differentiated P19 cells. (A) Western blots showing the levels of phosphorylated JNK in differentiated P19 cells treated with the Mn (0.3 mM), glutamate (G; 5 mM) and /or riluzole (R; 10 μM) for 18 hrs.; GAPDH antibody was used as a loading control. (B) Graphical representation of band densities from four separate experiments. Results are presented as mean ± SE; Mn vs. control a - p

    Article Snippet: Phospho-JNK antibody, anti-GAPDH antibody and secondary antibody for Western blots were obtained from Cell Signaling (Danvers, MA) and Western lightning plus substrate to develop immunoblots was from Perkin Elmer (Waltham, MA).

    Techniques: Western Blot

    miR-130b-3p participates in the regulation of Dsg1 expression. ( a ) Primary human keratinocytes were isolated as described in Materials and Methods and plated on dishes. Before they reached confluence, cells were induced to differentiate by adding 1.8 mM CaCl 2 to the culture medium. Cells were collected at the indicated time points to perform western blot analysis. Differentiation was evaluated by western blot for involucrin, K10 and TG1 ( a ). Dsg1 protein ( b ) upregulation during differentiation is also shown; GAPDH is used as loading control. The histograms show the quantitative mRNA level of involucrin, K10, TG1 and Dsg1 (fold over control; black bars) as mean±S.D. from three independent experiments. ( c ) Keratinocytes were exposed to calcium stimulation and miR-130b-3p levels were analyzed by qRT-PCR. Data are shown as mean±S.D. from three independent experiments. ( d ) Putative miR-130b-3p-binding site in the 3’-UTR region of Dsg1. ( e ) miR-130b-3p suppresses the expression of Dsg1. Keratinocytes were transfected with mimic-miR-130b-3p (miR-130b-3p) or mimic negative control (mimic-NC). The levels of Dsg1 were analyzed by immunoblot. n =3. ( f ) Knockdown of miR-130b-3p increases the expression levels of Dsg1. Keratinocytes were transfected with miR-130b-3p antagomir (anta-130b-3p) or the antagomir-negative control (anta-NC), the levels of Dsg1 were analyzed by immunoblot; n =3. ( g ) Keratinocytes were treated with miR-130b-3p or mimic-NC, and cells were exposed to 1.8 mM CaCl 2 for 72 h. The levels of Dsg1 were analyzed by immunoblot. n =3. * P

    Journal: Cell Death & Disease

    Article Title: H19 lncRNA regulates keratinocyte differentiation by targeting miR-130b-3p

    doi: 10.1038/cddis.2017.516

    Figure Lengend Snippet: miR-130b-3p participates in the regulation of Dsg1 expression. ( a ) Primary human keratinocytes were isolated as described in Materials and Methods and plated on dishes. Before they reached confluence, cells were induced to differentiate by adding 1.8 mM CaCl 2 to the culture medium. Cells were collected at the indicated time points to perform western blot analysis. Differentiation was evaluated by western blot for involucrin, K10 and TG1 ( a ). Dsg1 protein ( b ) upregulation during differentiation is also shown; GAPDH is used as loading control. The histograms show the quantitative mRNA level of involucrin, K10, TG1 and Dsg1 (fold over control; black bars) as mean±S.D. from three independent experiments. ( c ) Keratinocytes were exposed to calcium stimulation and miR-130b-3p levels were analyzed by qRT-PCR. Data are shown as mean±S.D. from three independent experiments. ( d ) Putative miR-130b-3p-binding site in the 3’-UTR region of Dsg1. ( e ) miR-130b-3p suppresses the expression of Dsg1. Keratinocytes were transfected with mimic-miR-130b-3p (miR-130b-3p) or mimic negative control (mimic-NC). The levels of Dsg1 were analyzed by immunoblot. n =3. ( f ) Knockdown of miR-130b-3p increases the expression levels of Dsg1. Keratinocytes were transfected with miR-130b-3p antagomir (anta-130b-3p) or the antagomir-negative control (anta-NC), the levels of Dsg1 were analyzed by immunoblot; n =3. ( g ) Keratinocytes were treated with miR-130b-3p or mimic-NC, and cells were exposed to 1.8 mM CaCl 2 for 72 h. The levels of Dsg1 were analyzed by immunoblot. n =3. * P

    Article Snippet: The anti-Dsg1 antibody (1:500, Abcam, Cambridge, MA, USA), anti-involucrin antibody (1:300, Abcam), anti-K10 antibody (1:500, Abcam), anti-TG1 (1:200, Santa Cruz Biotechnology CA, USA) and anti-GAPDH antibody (1:2,000, Abcam) were used in this study.

    Techniques: Expressing, Isolation, Western Blot, Quantitative RT-PCR, Binding Assay, Transfection, Negative Control

    Trends of actinomycin V on p53, p21 Waf1/Cip1 and Bax expression. ( A ) Expressions of relative proteins were measured by Western blotting; ( B ) Relative mRNA expression values were calculated by real-time PCR after 1 nmol/L actinomycin V treatment. The quantity of each mRNA was relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA levels. *** p

    Journal: Marine Drugs

    Article Title: Actinomycin V Suppresses Human Non-Small-Cell Lung Carcinoma A549 Cells by Inducing G2/M Phase Arrest and Apoptosis via the p53-Dependent Pathway

    doi: 10.3390/md17100572

    Figure Lengend Snippet: Trends of actinomycin V on p53, p21 Waf1/Cip1 and Bax expression. ( A ) Expressions of relative proteins were measured by Western blotting; ( B ) Relative mRNA expression values were calculated by real-time PCR after 1 nmol/L actinomycin V treatment. The quantity of each mRNA was relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA levels. *** p

    Article Snippet: Bcl-2, Bax and GAPDH antibodies were supplied by Abcam, Inc. (Cambridge, MA, USA).

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

    Evaluation of OmpA expression on A. hydrophila cells upon iron-deficient serum stimulation and bacterial susceptibility to complement attack. ( A , B , and C ) The steady state level of OmpA mRNA or the amount of OmpA protein in A. hydrophila cells cultured in the presence of unstimulated serum (Unstimulated), A. hydrophila DNA (1 μg per fish) stimulated serum plus 10 μM Fe 2+ supplemented ones (Iron added), and A. hydrophila DNA stimulated serum (Stimulated) was detected by Q-PCR ( A ), Western blot ( B ) and ELISA ( C ). The gray value of the relative signal intensity (OmpA/GAPDH) in Western blot analysis ( B ) was calculated using ImageJ program and depicted in a bar graph. All data are representative of at least three independent experiments. ( D ) Evaluation of the log 10 CFU of 100 μL A. hydrophila under the attack of guinea pig serum (diluted to 5% as the complement source). A. hydrophila cultured in the presence of differently treated serum samples followed by co-incubation with guinea pig serum. The green round spots (Unstimulated) showed Log 10 CFU of 100 μL A. hydrophila incubated with normal serum (n = 6), the red triangles (Iron added) showed Log 10 CFU of 100 μL A. hydrophila incubated with A. hydrophila DNA stimulated serum added with 10 μM iron (n = 6), and the blue squares (Stimulated) showed Log 10 CFU of 100 μL A. hydrophila incubated with A. hydrophila DNA stimulated serum (n = 6). The bars in the middle of each group mean average value. The steady state level of OmpA mRNA in A . hydrophila cells was calculated by 2 −ΔΔCT method normalized to 16 s rRNA. *p

    Journal: Scientific Reports

    Article Title: Coordination of Bactericidal and Iron Regulatory Functions of Hepcidin in Innate Antimicrobial Immunity in a Zebrafish Model

    doi: 10.1038/s41598-017-04069-x

    Figure Lengend Snippet: Evaluation of OmpA expression on A. hydrophila cells upon iron-deficient serum stimulation and bacterial susceptibility to complement attack. ( A , B , and C ) The steady state level of OmpA mRNA or the amount of OmpA protein in A. hydrophila cells cultured in the presence of unstimulated serum (Unstimulated), A. hydrophila DNA (1 μg per fish) stimulated serum plus 10 μM Fe 2+ supplemented ones (Iron added), and A. hydrophila DNA stimulated serum (Stimulated) was detected by Q-PCR ( A ), Western blot ( B ) and ELISA ( C ). The gray value of the relative signal intensity (OmpA/GAPDH) in Western blot analysis ( B ) was calculated using ImageJ program and depicted in a bar graph. All data are representative of at least three independent experiments. ( D ) Evaluation of the log 10 CFU of 100 μL A. hydrophila under the attack of guinea pig serum (diluted to 5% as the complement source). A. hydrophila cultured in the presence of differently treated serum samples followed by co-incubation with guinea pig serum. The green round spots (Unstimulated) showed Log 10 CFU of 100 μL A. hydrophila incubated with normal serum (n = 6), the red triangles (Iron added) showed Log 10 CFU of 100 μL A. hydrophila incubated with A. hydrophila DNA stimulated serum added with 10 μM iron (n = 6), and the blue squares (Stimulated) showed Log 10 CFU of 100 μL A. hydrophila incubated with A. hydrophila DNA stimulated serum (n = 6). The bars in the middle of each group mean average value. The steady state level of OmpA mRNA in A . hydrophila cells was calculated by 2 −ΔΔCT method normalized to 16 s rRNA. *p

    Article Snippet: After washing with TBST three times for 30 min, the blots were incubated with rabbit anti-hepcidin Ab (1:1,000), rabbit anti-OmpA Ab (1:1,000) or rabbit anti-GAPDH Ab (Abcam.

    Techniques: Expressing, Cell Culture, Fluorescence In Situ Hybridization, Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Incubation

    Evaluation of the induced expression of hepcidin and reduced serum iron concentration upon A. hydrophila DNA stimulation. ( A ) Detection of the steady state level of hepcidin mRNA in liver upon stimulation with various concentrations of A. hydrophila DNA (0.5, 1, 3, and 5 μg per fish) by Q-PCR. ( B and C ) Detection of the hepcidin protein level in serum upon stimulation with various concentrations of A. hydrophila DNA (0.5, 1, 3, and 5 μg per fish) by Western blot ( B ) and ELISA ( C ), and the gray value of the relative signal intensity (hepcidin/GAPDH) in Western blot analysis was calculated using ImageJ program and depicted in a bar graph. ( D ) Time-dependent upregulation of the steady state level of hepcidin mRNA in liver upon stimulation with A. hydrophila DNA (1 μg per fish) as determined by Q-PCR. ( E ) Time-dependent upregulation of the hepcidin protein in serum upon stimulation with A. hydrophila DNA (1 μg per fish) as determined by ELISA. ( F ) Reduced concentrations of serum iron stimulated by A. hydrophila DNA (0.5, 1, 3, and 5 μg per fish) were detected by ICP-MS. The control groups in each experiment (indicated as Control) received mock PBS. The steady state level of hepcidin mRNA in liver was calculated by 2 −ΔΔCT method normalized to β-actin. All data are representative of at least three independent experiments. A significant difference was detected between each experimental and control group. *p

    Journal: Scientific Reports

    Article Title: Coordination of Bactericidal and Iron Regulatory Functions of Hepcidin in Innate Antimicrobial Immunity in a Zebrafish Model

    doi: 10.1038/s41598-017-04069-x

    Figure Lengend Snippet: Evaluation of the induced expression of hepcidin and reduced serum iron concentration upon A. hydrophila DNA stimulation. ( A ) Detection of the steady state level of hepcidin mRNA in liver upon stimulation with various concentrations of A. hydrophila DNA (0.5, 1, 3, and 5 μg per fish) by Q-PCR. ( B and C ) Detection of the hepcidin protein level in serum upon stimulation with various concentrations of A. hydrophila DNA (0.5, 1, 3, and 5 μg per fish) by Western blot ( B ) and ELISA ( C ), and the gray value of the relative signal intensity (hepcidin/GAPDH) in Western blot analysis was calculated using ImageJ program and depicted in a bar graph. ( D ) Time-dependent upregulation of the steady state level of hepcidin mRNA in liver upon stimulation with A. hydrophila DNA (1 μg per fish) as determined by Q-PCR. ( E ) Time-dependent upregulation of the hepcidin protein in serum upon stimulation with A. hydrophila DNA (1 μg per fish) as determined by ELISA. ( F ) Reduced concentrations of serum iron stimulated by A. hydrophila DNA (0.5, 1, 3, and 5 μg per fish) were detected by ICP-MS. The control groups in each experiment (indicated as Control) received mock PBS. The steady state level of hepcidin mRNA in liver was calculated by 2 −ΔΔCT method normalized to β-actin. All data are representative of at least three independent experiments. A significant difference was detected between each experimental and control group. *p

    Article Snippet: After washing with TBST three times for 30 min, the blots were incubated with rabbit anti-hepcidin Ab (1:1,000), rabbit anti-OmpA Ab (1:1,000) or rabbit anti-GAPDH Ab (Abcam.

    Techniques: Expressing, Concentration Assay, Fluorescence In Situ Hybridization, Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay, Mass Spectrometry