anti beta actin  (Millipore)


Bioz Verified Symbol Millipore is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    Anti beta Actin antibody
    Description:
    The ACTB β actin gene is mapped to human chromosome 7p22 1 β Actin is the most abundant protein localized to the cytoplasm ACTB is expressed ubiquitously Actin is one of the most conserved eukaryotic proteins it is expressed in mammals and birds Four of the actin isoforms represent the differentiation markers of muscle tissues and two are found in almost all cells Anti β Actin antibody Mouse Monoclonal mouse IgG1 isotype is derived from the AC 15 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from BALB c mice immunized with a slightly modified synthetic b cytoplasmic actin N terminal peptide Ac Asp Asp Asp Ile Ala Ala Leu Val Ile Asp Asn Gly Ser Gly Lys conjugated to KLH In staining of chicken gizzard ultrathin tissue cryosections the antibody labels the dense bodies and longitudinal channels linking consecutive dense bodies that are also occupied by desmin and the membrane associated dense plaque It does not stain adult cardiac and skeletal muscles except for traces due to contaminations of the sample with non muscle cells or if embryonic tissue is being used
    Catalog Number:
    a1978
    Price:
    None
    Applications:
    The actin in cells of various species and tissue origin is very similar in their immunological and physical properties. As a consequence, it has been difficult to produce potent antisera to this protein. Therefore the availability of monoclonal antibodies to β-actin provides a specific and useful tool in studying the intracellular distribution of β-actin and the static and dynamic aspects of the cytoskeleton.Monoclonal anti-beta-actin antibody can be used for microarray, indirect immunofluorescence, and immunohistochemical analyses. Furthermore, the product has been used for immunocytochemistry at 10-40 μg/mL using human foreskin fibroblasts. The antibody has also been used for western blot at 0.5-1 μg/mL using cell extract of human foreskin fibroblasts or chicken fibroblasts.The antibody can be used for staining of acetone-fixed frozen sections, EM preparations, and microinjection experiments. B5, ethanol, methacam, or Bouin′s solutions can be used as fixatives. The epitope recognized by the antibody is resistant to formalin-fixation and paraffin-embedding.It has been used in Immunoblot analysis.
    Buy from Supplier


    Structured Review

    Millipore anti beta actin
    Anti beta Actin antibody
    The ACTB β actin gene is mapped to human chromosome 7p22 1 β Actin is the most abundant protein localized to the cytoplasm ACTB is expressed ubiquitously Actin is one of the most conserved eukaryotic proteins it is expressed in mammals and birds Four of the actin isoforms represent the differentiation markers of muscle tissues and two are found in almost all cells Anti β Actin antibody Mouse Monoclonal mouse IgG1 isotype is derived from the AC 15 hybridoma produced by the fusion of mouse myeloma cells and splenocytes from BALB c mice immunized with a slightly modified synthetic b cytoplasmic actin N terminal peptide Ac Asp Asp Asp Ile Ala Ala Leu Val Ile Asp Asn Gly Ser Gly Lys conjugated to KLH In staining of chicken gizzard ultrathin tissue cryosections the antibody labels the dense bodies and longitudinal channels linking consecutive dense bodies that are also occupied by desmin and the membrane associated dense plaque It does not stain adult cardiac and skeletal muscles except for traces due to contaminations of the sample with non muscle cells or if embryonic tissue is being used
    https://www.bioz.com/result/anti beta actin/product/Millipore
    Average 99 stars, based on 28 article reviews
    Price from $9.99 to $1999.99
    anti beta actin - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Nuclear import of Xenopus egg extract components into cultured cells for reprogramming purposes: a case study on goldfish fin cells"

    Article Title: Nuclear import of Xenopus egg extract components into cultured cells for reprogramming purposes: a case study on goldfish fin cells

    Journal: Scientific Reports

    doi: 10.1038/s41598-019-39500-y

    Nuclear import of the fusion protein GST-GFP-NLS in the permeabilized cells. Cells permeabilized with 30 µg/mL digitonin (Dig30) and non-permeabilized cells (Control) were incubated with GST-GFP-NLS in the absence (−Extract) or presence (+Extract) of Xenopus egg extract supplemented with ATP for 1 h at 25 °C. The incorporation of GST-GFP-NLS was assessed by green fluorescence. Cell nuclei were counterstained with Hoechst 33243. Note that without egg extract, the nuclei are not labelled (arrows). Due to light scattering of the fluorescence in the cytoplasm, the nuclei appear smaller than they are. The egg extract restored nuclear import in the permeabilized cells. These pictures are representative of five independent replicates. Scale bar = 20 µm. Detection of importin alpha-1 (55 kDa) ( B ) and Karyopherin beta -1 (KPNB1–97 kDa) ( C ) in Xenopus egg extract by western blot in the presence (+) or in absence (−) of the anti- Xenopus importin alpha-1 antibody (clone 15) and the anti-rat KPNB1 antibody (KPNB1-clone 23). Beta actin was used as a loading control. M: size markers. The cropped blots came from the same gels and were analyzed with the same exposure times (B:1 sec; C:1 min).
    Figure Legend Snippet: Nuclear import of the fusion protein GST-GFP-NLS in the permeabilized cells. Cells permeabilized with 30 µg/mL digitonin (Dig30) and non-permeabilized cells (Control) were incubated with GST-GFP-NLS in the absence (−Extract) or presence (+Extract) of Xenopus egg extract supplemented with ATP for 1 h at 25 °C. The incorporation of GST-GFP-NLS was assessed by green fluorescence. Cell nuclei were counterstained with Hoechst 33243. Note that without egg extract, the nuclei are not labelled (arrows). Due to light scattering of the fluorescence in the cytoplasm, the nuclei appear smaller than they are. The egg extract restored nuclear import in the permeabilized cells. These pictures are representative of five independent replicates. Scale bar = 20 µm. Detection of importin alpha-1 (55 kDa) ( B ) and Karyopherin beta -1 (KPNB1–97 kDa) ( C ) in Xenopus egg extract by western blot in the presence (+) or in absence (−) of the anti- Xenopus importin alpha-1 antibody (clone 15) and the anti-rat KPNB1 antibody (KPNB1-clone 23). Beta actin was used as a loading control. M: size markers. The cropped blots came from the same gels and were analyzed with the same exposure times (B:1 sec; C:1 min).

    Techniques Used: Incubation, Fluorescence, Western Blot, Size-exclusion Chromatography

    Incorporation of Lamin B3 from Xenopus eggs into the nuclei of permeabilized cells. ( A ) Detection of Lamin B3 (68 kDa) in Xenopus egg extract (Extract) by western blot in the presence (+) or absence (−) of anti- Xenopus Lamin B3. Beta actin was used as a loading control. M: size markers. The cropped blots came from the same gel and were analyzed with the same exposure times (Lamin B3:2 sec; beta actin: 1 min). ( B ) Nuclear import of Lamin B3 detected by immunofluorescence in permeabilized (Dig30) and non-permeabilized (Control) cells incubated for 60 min in the presence (+Extract) of Xenopus egg extract under energy supplementation at 25 °C. Lamin B3-positive cells presenting a detectable signal (from weak to strong staining) were observed. A high proportion of nuclei were labelled. Inset: magnification of one nucleus showing a strong labelling of the nuclear lamina. These pictures are representative of five independent replicates. Scale bar = 20 µm.
    Figure Legend Snippet: Incorporation of Lamin B3 from Xenopus eggs into the nuclei of permeabilized cells. ( A ) Detection of Lamin B3 (68 kDa) in Xenopus egg extract (Extract) by western blot in the presence (+) or absence (−) of anti- Xenopus Lamin B3. Beta actin was used as a loading control. M: size markers. The cropped blots came from the same gel and were analyzed with the same exposure times (Lamin B3:2 sec; beta actin: 1 min). ( B ) Nuclear import of Lamin B3 detected by immunofluorescence in permeabilized (Dig30) and non-permeabilized (Control) cells incubated for 60 min in the presence (+Extract) of Xenopus egg extract under energy supplementation at 25 °C. Lamin B3-positive cells presenting a detectable signal (from weak to strong staining) were observed. A high proportion of nuclei were labelled. Inset: magnification of one nucleus showing a strong labelling of the nuclear lamina. These pictures are representative of five independent replicates. Scale bar = 20 µm.

    Techniques Used: Western Blot, Size-exclusion Chromatography, Immunofluorescence, Incubation, Staining

    2) Product Images from "Interaction of septin 7 and DOCK8 in equine lymphocytes reveals novel insights into signaling pathways associated with autoimmunity"

    Article Title: Interaction of septin 7 and DOCK8 in equine lymphocytes reveals novel insights into signaling pathways associated with autoimmunity

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-30753-7

    Diminished DOCK8 protein levels in PBL of ERU cases. (A) Representative signal abundances of DOCK8 in PBL of controls (left panel) and ERU (right panel) detected by western blot. DOCK8 signal abundance in PBL of diseased specimen was decreased compared to controls. Lower molecular extra bands result from further reactivities of the polyclonal rabbit anti DOCK8 antibody. (B) Compared to healthy controls (n = 16), DOCK8 levels in ERU PBL (n = 36) were significantly (***p ≤ 0.001) decreased to 63% of physiological expression rate (set to a 100%). All protein abundances of DOCK8 were normalized to beta actin. (C) Analyzes of lymphocyte subset composition in healthy controls and autoimmune cases revealed no significant differences. (D) Statistical analysis of DOCK8 expression differences in lymphocyte subsets of healthy controls (white bars; n = 13) and ERU cases (grey bars; n = 12). DOCK8 expression intensity (geo MFI) was decreased in all lymphocyte subsets of ERU cases. In CD8 + T cells, no considerable DOCK8 expression difference between control PBL and ERU cases could be detected, whereas in CD4 + T cells of ERU horses, geo MFI was reduced to 85% compared to healthy controls. In B cells of autoimmune cases, a significant decrease of DOCK8 geo MFI to 57% was detected (*p ≤ 0.05).
    Figure Legend Snippet: Diminished DOCK8 protein levels in PBL of ERU cases. (A) Representative signal abundances of DOCK8 in PBL of controls (left panel) and ERU (right panel) detected by western blot. DOCK8 signal abundance in PBL of diseased specimen was decreased compared to controls. Lower molecular extra bands result from further reactivities of the polyclonal rabbit anti DOCK8 antibody. (B) Compared to healthy controls (n = 16), DOCK8 levels in ERU PBL (n = 36) were significantly (***p ≤ 0.001) decreased to 63% of physiological expression rate (set to a 100%). All protein abundances of DOCK8 were normalized to beta actin. (C) Analyzes of lymphocyte subset composition in healthy controls and autoimmune cases revealed no significant differences. (D) Statistical analysis of DOCK8 expression differences in lymphocyte subsets of healthy controls (white bars; n = 13) and ERU cases (grey bars; n = 12). DOCK8 expression intensity (geo MFI) was decreased in all lymphocyte subsets of ERU cases. In CD8 + T cells, no considerable DOCK8 expression difference between control PBL and ERU cases could be detected, whereas in CD4 + T cells of ERU horses, geo MFI was reduced to 85% compared to healthy controls. In B cells of autoimmune cases, a significant decrease of DOCK8 geo MFI to 57% was detected (*p ≤ 0.05).

    Techniques Used: Western Blot, Expressing

    ILK expression in lymphocytes was significantly increased in diseased state and coincidented with increased cell survival. (A) Significantly increased ILK protein levels in ERU cases (n = 33) compared to healthy controls (n = 18) were verified and quantified by western blots (**p ≤ 0.01). ILK intensities were normalized to beta actin abundances on respective blots. Representative signal abundances are shown above respective columns. (B) PBL of healthy (biological n = 4, technical replicates n = 2) and ERU horses (biological replicates n = 6), which constitutively differed in ILK expression were cultured at 37 °C for 72 hours. Apoptosis rate was determined by measuring the amount of annexin V positive cells in flow cytometry. ERU horses (grey column), expressing ILK significantly higher (1.5-fold), showed a highly significantly reduced amount of apoptotic cells compared to healthy controls (white column) (**p ≤ 0.01). (C) In contrast, necrosis rate of lymphocytes, determined by the amount of PI positive cells, was not different between healthy controls (white column) and ERU cases (grey bar).
    Figure Legend Snippet: ILK expression in lymphocytes was significantly increased in diseased state and coincidented with increased cell survival. (A) Significantly increased ILK protein levels in ERU cases (n = 33) compared to healthy controls (n = 18) were verified and quantified by western blots (**p ≤ 0.01). ILK intensities were normalized to beta actin abundances on respective blots. Representative signal abundances are shown above respective columns. (B) PBL of healthy (biological n = 4, technical replicates n = 2) and ERU horses (biological replicates n = 6), which constitutively differed in ILK expression were cultured at 37 °C for 72 hours. Apoptosis rate was determined by measuring the amount of annexin V positive cells in flow cytometry. ERU horses (grey column), expressing ILK significantly higher (1.5-fold), showed a highly significantly reduced amount of apoptotic cells compared to healthy controls (white column) (**p ≤ 0.01). (C) In contrast, necrosis rate of lymphocytes, determined by the amount of PI positive cells, was not different between healthy controls (white column) and ERU cases (grey bar).

    Techniques Used: Expressing, Western Blot, Cell Culture, Flow Cytometry, Cytometry

    3) Product Images from "Distinct and overlapping gene regulatory networks in BMP- and HDAC-controlled cell fate determination in the embryonic forebrain"

    Article Title: Distinct and overlapping gene regulatory networks in BMP- and HDAC-controlled cell fate determination in the embryonic forebrain

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-13-298

    Quantitative real-time PCR. Total RNA was extracted from TSA and BMP2 treated neurosphere cultures. Neurosphere cultures were cultured for 7 days and subsequently dissociated and plated out in monolayers. Cells were treated with TSA (10, 25, 50nM) or 10 ng/μl BMP2 from 1.5-2.5 days after plating. Total RNA was collected 6, 12 or 24 h upon treatment. Reverse-transcript cDNA was analyzed using primer recognizing the following genes: Bmp2 ( A ), Bmp4 ( B ), Id1 ( C ), Id2 ( D ), Smad7 ( E ), Stat3 ( F ). Gpr17 ( G ), Bambi ( H ), Wnt5a ( I ), Wisp1 ( J ) and Primer recognizing beta-Actin and Gapdh were used for normalization. Shown values represent the mean (+/− SEM) of three individual experiments (6 h and 24 h) or two individual experiments (12 h). * = p
    Figure Legend Snippet: Quantitative real-time PCR. Total RNA was extracted from TSA and BMP2 treated neurosphere cultures. Neurosphere cultures were cultured for 7 days and subsequently dissociated and plated out in monolayers. Cells were treated with TSA (10, 25, 50nM) or 10 ng/μl BMP2 from 1.5-2.5 days after plating. Total RNA was collected 6, 12 or 24 h upon treatment. Reverse-transcript cDNA was analyzed using primer recognizing the following genes: Bmp2 ( A ), Bmp4 ( B ), Id1 ( C ), Id2 ( D ), Smad7 ( E ), Stat3 ( F ). Gpr17 ( G ), Bambi ( H ), Wnt5a ( I ), Wisp1 ( J ) and Primer recognizing beta-Actin and Gapdh were used for normalization. Shown values represent the mean (+/− SEM) of three individual experiments (6 h and 24 h) or two individual experiments (12 h). * = p

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

    Western Blot and ELISA Microarray. Proteins were extracted from TSA (10, 25, 50nM) and BMP2 (10 ng/μl) treated neurosphere cultures 6, 12 or 24 h after treatment. Western Blot for pSmad1/5/8, Smad1/5/8, pStat3, Stat3, pGsk3-beta was performed. Beta-Actin was used as control ( A ). Phosphorylated proteins were quantified using an ArrayTube™ (Alere Technologies, Jena, Germany) based sandwich ELISA microarray. Concentration of pGsk3-beta ( B ) and pErk2 ( C ) in relation to total protein content is presented.
    Figure Legend Snippet: Western Blot and ELISA Microarray. Proteins were extracted from TSA (10, 25, 50nM) and BMP2 (10 ng/μl) treated neurosphere cultures 6, 12 or 24 h after treatment. Western Blot for pSmad1/5/8, Smad1/5/8, pStat3, Stat3, pGsk3-beta was performed. Beta-Actin was used as control ( A ). Phosphorylated proteins were quantified using an ArrayTube™ (Alere Technologies, Jena, Germany) based sandwich ELISA microarray. Concentration of pGsk3-beta ( B ) and pErk2 ( C ) in relation to total protein content is presented.

    Techniques Used: Western Blot, Enzyme-linked Immunosorbent Assay, Microarray, Sandwich ELISA, Concentration Assay

    4) Product Images from "DJ-1 Knockout Augments Disease Severity and Shortens Survival in a Mouse Model of ALS"

    Article Title: DJ-1 Knockout Augments Disease Severity and Shortens Survival in a Mouse Model of ALS

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0117190

    Motor neuron loss and astrogliosis in the spinal cords of SOD1 DJ-1 KO. Choline acyl transferase (ChAT, A) and glial fibrillary acidic protein (GFAP, B) levels were determined in the spinal cord extracts by Western blot analysis. Proteins levels were normalized to beta actin. Data is presented as averages ± SD. * p
    Figure Legend Snippet: Motor neuron loss and astrogliosis in the spinal cords of SOD1 DJ-1 KO. Choline acyl transferase (ChAT, A) and glial fibrillary acidic protein (GFAP, B) levels were determined in the spinal cord extracts by Western blot analysis. Proteins levels were normalized to beta actin. Data is presented as averages ± SD. * p

    Techniques Used: Western Blot

    Attenuated Nrf2 system in SOD DJ-1 KO mice. Mutated SOD1 increased the expression of Nrf2 and HO-1 while SOD1 DJ-1 KO demonstrated reduction. Nuclear factor erythroid-2 related factor 2 (Nrf2, A) and hemeoxygenase 1(HO-1, B) protein levels were determined in the spinal cord extracts by Western blot anlysis. Proteins levels were normalized to beta actin. Data is presented as averages ± SD. * p
    Figure Legend Snippet: Attenuated Nrf2 system in SOD DJ-1 KO mice. Mutated SOD1 increased the expression of Nrf2 and HO-1 while SOD1 DJ-1 KO demonstrated reduction. Nuclear factor erythroid-2 related factor 2 (Nrf2, A) and hemeoxygenase 1(HO-1, B) protein levels were determined in the spinal cord extracts by Western blot anlysis. Proteins levels were normalized to beta actin. Data is presented as averages ± SD. * p

    Techniques Used: Mouse Assay, Expressing, Western Blot

    5) Product Images from "PGRMC1 phosphorylation affects cell shape, motility, glycolysis, mitochondrial form and function, and tumor growth"

    Article Title: PGRMC1 phosphorylation affects cell shape, motility, glycolysis, mitochondrial form and function, and tumor growth

    Journal: BMC Molecular and Cell Biology

    doi: 10.1186/s12860-020-00256-3

    MIA PaCa-2 pancreatic cancer cells morphology is affected by PGRMC1 phosphorylation status. a PGRMC1-HA proteins constructed for this figure. TMH: Trans-membrane helix. HA: the C-terminal 3x hemaglutinin tag. b Detection of exogenous PGRMC1 expression levels by western blot (upper panel). Equal loading is controlled by quantifying beta actin (lower panel). The results show three totally independent stably transfected cell lines per plasmid from (A). Open arrow: Exogenous PGRMC1-HA (Ex.). Shaded arrow: endogenous PGRMC1 (End.). Filled arrow: beta actin. The molecular weight ladder is Bio-Rad 1610377 Dual Xtra Standards. c Box plots quantification of replicate gels of (B) with signals normalized to beta actin from the same respective lanes. n = 4 lanes for MP and n = 6 for WT, DM and TM (replicates of respective lines 1–3 per condition). There were no significant differences (ns) except for the exogenous band in MP (ANOVA, post-hoc Dunnet’s T3). d Western blot quantification of HA-tagged exogenous PGRMC1, following B but detecting PGRMC1 with anti-HA antibody. The molecular weight ladder is Abcam ab116028 Prestained Protein Ladder. e PGRMC1 mutant protein expression alters MIA PaCa-2 cell morphology. PGRMC1-HA-expressing stable cells (respective lines 1 from B) or MP cells were stained with a FITC-tagged anti-HA antibody (Anti-HA) and imaged by confocal microscopy. DNA was stained with DAPI. Cells were also imaged in differential interference contrast (DIC) microscopy mode. The respective left panels show merged images of all 3 channels. f The rounded phenotype of double and triple mutant (E) was reversed to elongated phenotype after 125 μM ROCKI addition, but not by addition of DMSO vehicle control
    Figure Legend Snippet: MIA PaCa-2 pancreatic cancer cells morphology is affected by PGRMC1 phosphorylation status. a PGRMC1-HA proteins constructed for this figure. TMH: Trans-membrane helix. HA: the C-terminal 3x hemaglutinin tag. b Detection of exogenous PGRMC1 expression levels by western blot (upper panel). Equal loading is controlled by quantifying beta actin (lower panel). The results show three totally independent stably transfected cell lines per plasmid from (A). Open arrow: Exogenous PGRMC1-HA (Ex.). Shaded arrow: endogenous PGRMC1 (End.). Filled arrow: beta actin. The molecular weight ladder is Bio-Rad 1610377 Dual Xtra Standards. c Box plots quantification of replicate gels of (B) with signals normalized to beta actin from the same respective lanes. n = 4 lanes for MP and n = 6 for WT, DM and TM (replicates of respective lines 1–3 per condition). There were no significant differences (ns) except for the exogenous band in MP (ANOVA, post-hoc Dunnet’s T3). d Western blot quantification of HA-tagged exogenous PGRMC1, following B but detecting PGRMC1 with anti-HA antibody. The molecular weight ladder is Abcam ab116028 Prestained Protein Ladder. e PGRMC1 mutant protein expression alters MIA PaCa-2 cell morphology. PGRMC1-HA-expressing stable cells (respective lines 1 from B) or MP cells were stained with a FITC-tagged anti-HA antibody (Anti-HA) and imaged by confocal microscopy. DNA was stained with DAPI. Cells were also imaged in differential interference contrast (DIC) microscopy mode. The respective left panels show merged images of all 3 channels. f The rounded phenotype of double and triple mutant (E) was reversed to elongated phenotype after 125 μM ROCKI addition, but not by addition of DMSO vehicle control

    Techniques Used: Construct, Expressing, Western Blot, Stable Transfection, Transfection, Plasmid Preparation, Molecular Weight, Mutagenesis, Staining, Confocal Microscopy, Microscopy

    6) Product Images from "Betaine Alleviates Hypertriglycemia and Tau Hyperphosphorylation in db/db Mice"

    Article Title: Betaine Alleviates Hypertriglycemia and Tau Hyperphosphorylation in db/db Mice

    Journal: Toxicological Research

    doi: 10.5487/TR.2013.29.1.007

    Effect of betaine supplementation on glucose and lipid metabolism in the livers of db/db mice. (A) Relative mRNA levels of PGC-1α were determined by semiquantitative RT-PCR. Equal RNA loading was confirmed with beta-actin. (B) Relative mRNA levels of PPARα, CPT1a, and SCD1 were determined by real-time quantitative RT-PCR (n = 3-4). (C) The protein levels of p-ACC and beta-actin were determined by immunoblotting analysis (n = 3). Each bar represents the mean ± SEM. * p
    Figure Legend Snippet: Effect of betaine supplementation on glucose and lipid metabolism in the livers of db/db mice. (A) Relative mRNA levels of PGC-1α were determined by semiquantitative RT-PCR. Equal RNA loading was confirmed with beta-actin. (B) Relative mRNA levels of PPARα, CPT1a, and SCD1 were determined by real-time quantitative RT-PCR (n = 3-4). (C) The protein levels of p-ACC and beta-actin were determined by immunoblotting analysis (n = 3). Each bar represents the mean ± SEM. * p

    Techniques Used: Mouse Assay, Pyrolysis Gas Chromatography, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

    (A) Effect of betaine supplementation on ER stress marker in the brains of db/db mice. Protein levels of BiP and beta-actin were determined by immunoblotting analysis. (B) Effect of betaine supplementation on JNK activation in the brains of db/db mice. The protein levels of p-JNK and total JNK3 were determined by immunoblotting analysis. (C) Effect of betaine supplementation on tau hyperphosphorylation in the brains of db/db mice. The protein levels of dephosphorylated tau were determined by immunoblotting analysis.
    Figure Legend Snippet: (A) Effect of betaine supplementation on ER stress marker in the brains of db/db mice. Protein levels of BiP and beta-actin were determined by immunoblotting analysis. (B) Effect of betaine supplementation on JNK activation in the brains of db/db mice. The protein levels of p-JNK and total JNK3 were determined by immunoblotting analysis. (C) Effect of betaine supplementation on tau hyperphosphorylation in the brains of db/db mice. The protein levels of dephosphorylated tau were determined by immunoblotting analysis.

    Techniques Used: Marker, Mouse Assay, Activation Assay

    7) Product Images from "Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6 zygotes"

    Article Title: Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6 zygotes

    Journal: BMC Biotechnology

    doi: 10.1186/s12896-016-0234-4

    Cas9 is expressed in B cells of Rosa26 LSL-Cas9 knock-in mice. a : Strategy for isolating naive B cells from spleens of three Rosa26 LSL-Cas9 F1 mice by CD43 depletion and activation of Cas9 expression by deletion of the loxP flanked stop element upon treatment with TAT-Cre protein. b : Scheme of the TAT-Cre-mediated deletion of the Neo/Stop element (left). Detection of Cre-mediated deletion of the Neo/STOP cassette by PCR using DNA of TAT-Cre/LPS-treated B cells and the indicated primers. Rosa26 LSL-Cas9 alleles produce a 1.0 kb band (CAGF-NeoR1 primers), Cre recombined alleles are detected by a 0.7 kb band (CAGF/Cas9R1 primers). c : Sequencing results of 0.7 kb PCR products (from b ) showing the correct deletion of the loxP flanked stop element, leaving one loxP site in between the CAG promoter and the Cas9 coding region. d : Western blot analysis of lysates prepared from TAT-Cre/LPS treated B cells of three Rosa26 LSL-Cas9 F1 mice using antibodies against the Flag Tag, Cas9, or Beta-actin
    Figure Legend Snippet: Cas9 is expressed in B cells of Rosa26 LSL-Cas9 knock-in mice. a : Strategy for isolating naive B cells from spleens of three Rosa26 LSL-Cas9 F1 mice by CD43 depletion and activation of Cas9 expression by deletion of the loxP flanked stop element upon treatment with TAT-Cre protein. b : Scheme of the TAT-Cre-mediated deletion of the Neo/Stop element (left). Detection of Cre-mediated deletion of the Neo/STOP cassette by PCR using DNA of TAT-Cre/LPS-treated B cells and the indicated primers. Rosa26 LSL-Cas9 alleles produce a 1.0 kb band (CAGF-NeoR1 primers), Cre recombined alleles are detected by a 0.7 kb band (CAGF/Cas9R1 primers). c : Sequencing results of 0.7 kb PCR products (from b ) showing the correct deletion of the loxP flanked stop element, leaving one loxP site in between the CAG promoter and the Cas9 coding region. d : Western blot analysis of lysates prepared from TAT-Cre/LPS treated B cells of three Rosa26 LSL-Cas9 F1 mice using antibodies against the Flag Tag, Cas9, or Beta-actin

    Techniques Used: Knock-In, Mouse Assay, Activation Assay, Expressing, Polymerase Chain Reaction, Sequencing, Western Blot, FLAG-tag

    8) Product Images from "Transgenic expression of the N525S-tuberin variant in Tsc2 mutant (Eker) rats causes dominant embryonic lethality"

    Article Title: Transgenic expression of the N525S-tuberin variant in Tsc2 mutant (Eker) rats causes dominant embryonic lethality

    Journal: Scientific Reports

    doi: 10.1038/srep05927

    Histological analysis of NSM Tg-positive embryos. (a) Macroscopic analysis of embryos. There were no apparent abnormalities or difference in size among individual embryos at any stage irrespective of their NSM Tg genetic status. (b) Class III beta-tubulin (TuJ1) immunohistochemical (IHC) analysis of embryonic tissue sections. Representative coronal sections of the embryo heads carrying NSM Tg are shown. Exencephaly, which is frequently associated with lethality in Eker homozygotes, was not observed.
    Figure Legend Snippet: Histological analysis of NSM Tg-positive embryos. (a) Macroscopic analysis of embryos. There were no apparent abnormalities or difference in size among individual embryos at any stage irrespective of their NSM Tg genetic status. (b) Class III beta-tubulin (TuJ1) immunohistochemical (IHC) analysis of embryonic tissue sections. Representative coronal sections of the embryo heads carrying NSM Tg are shown. Exencephaly, which is frequently associated with lethality in Eker homozygotes, was not observed.

    Techniques Used: Immunohistochemistry

    9) Product Images from "A Humanin Derivative Reduces Amyloid Beta Accumulation and Ameliorates Memory Deficit in Triple Transgenic Mice"

    Article Title: A Humanin Derivative Reduces Amyloid Beta Accumulation and Ameliorates Memory Deficit in Triple Transgenic Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0016259

    Effect of S14G-HN on tau phosphorylation. A, B. Brain homogenate of 3xTg-AD mice treated with S14G-HN (H) or vehicle (V) was subjected to immunoblot analysis using anti-phosphorylated tau (Ser202) (AT8) (upper panel in A), anti-phosphorylated tau (Thr231) (AT180) (upper panel in B), anti-total tau (tau46) (middle panels) and anti-beta-actin (lower panels) antibodies. C–F. Brain sections were subjected to immunostaining using anti-phosphorylated tau (Ser202) (AT8) antibody ( C, D ) or anti-phosphorylated tau (Thr231) (AT180) antibody ( E, F ) and biotin-conjugated secondary antibody followed by visualization with ABC method. Representative images of hippocampal CA1 region of vehicle ( C, E ) and S14G-HN ( D, F ) treated mouse sections are shown. Bar = 200 µm.
    Figure Legend Snippet: Effect of S14G-HN on tau phosphorylation. A, B. Brain homogenate of 3xTg-AD mice treated with S14G-HN (H) or vehicle (V) was subjected to immunoblot analysis using anti-phosphorylated tau (Ser202) (AT8) (upper panel in A), anti-phosphorylated tau (Thr231) (AT180) (upper panel in B), anti-total tau (tau46) (middle panels) and anti-beta-actin (lower panels) antibodies. C–F. Brain sections were subjected to immunostaining using anti-phosphorylated tau (Ser202) (AT8) antibody ( C, D ) or anti-phosphorylated tau (Thr231) (AT180) antibody ( E, F ) and biotin-conjugated secondary antibody followed by visualization with ABC method. Representative images of hippocampal CA1 region of vehicle ( C, E ) and S14G-HN ( D, F ) treated mouse sections are shown. Bar = 200 µm.

    Techniques Used: Mouse Assay, Immunostaining

    S14G-HN does not affect APP production and processing. Brain homogenate of 3xTg-AD mice treated with S14G-HN (HNG or H) or vehicle (veh or V) was subjected to immunoblot analysis using anti-APP C-terminus ( A ) or anti-sAPPalpha ( B ) (upper panels) and anti-beta-actin (lower panels) antibodies.
    Figure Legend Snippet: S14G-HN does not affect APP production and processing. Brain homogenate of 3xTg-AD mice treated with S14G-HN (HNG or H) or vehicle (veh or V) was subjected to immunoblot analysis using anti-APP C-terminus ( A ) or anti-sAPPalpha ( B ) (upper panels) and anti-beta-actin (lower panels) antibodies.

    Techniques Used: Mouse Assay

    10) Product Images from "Beta cell specific ZnT8 deletion in mice causes marked defects in insulin processing, crystallization and secretion"

    Article Title: Beta cell specific ZnT8 deletion in mice causes marked defects in insulin processing, crystallization and secretion

    Journal: Diabetologia

    doi: 10.1007/s00125-010-1733-9

    Role of ZnT8 and zinc in insulin biosynthesis and secretion in control and ZnT8BKO mouse beta cells. 1. Insulin transcription mediated via transcription factors such as PDX1 and MafA. 2. Proinsulin synthesis. 3. Zinc-proinsulin hexamerization. 4. ZnT8 is the primary zinc transporter on insulin granules and its loss in ZnT8BKO beta cells reduces zinc content. 5. Proinsulin to insulin conversion catalysed by PC1, PC2 and CpE enzymes. Enzyme expression is reduced in ZnT8BKO beta cells, increasing proinsulin content. 6. Generation of dense core granules following insulin crystallization. Reduced zinc-insulin crystallization, increased immature, abnormal and empty granules in ZnT8BKO beta cells was observed. 7. Release of insulin and zinc during granule exocytosis. ZnT8BKO beta cells show reduced insulin secretion and increased proinsulin secretion. ↑ increase, ↓ decrease, ⊥ inhibition
    Figure Legend Snippet: Role of ZnT8 and zinc in insulin biosynthesis and secretion in control and ZnT8BKO mouse beta cells. 1. Insulin transcription mediated via transcription factors such as PDX1 and MafA. 2. Proinsulin synthesis. 3. Zinc-proinsulin hexamerization. 4. ZnT8 is the primary zinc transporter on insulin granules and its loss in ZnT8BKO beta cells reduces zinc content. 5. Proinsulin to insulin conversion catalysed by PC1, PC2 and CpE enzymes. Enzyme expression is reduced in ZnT8BKO beta cells, increasing proinsulin content. 6. Generation of dense core granules following insulin crystallization. Reduced zinc-insulin crystallization, increased immature, abnormal and empty granules in ZnT8BKO beta cells was observed. 7. Release of insulin and zinc during granule exocytosis. ZnT8BKO beta cells show reduced insulin secretion and increased proinsulin secretion. ↑ increase, ↓ decrease, ⊥ inhibition

    Techniques Used: Expressing, Crystallization Assay, Inhibition

    Islet and beta cell morphological analysis in ZnT8BKO mice. a. Islet size (μm 2 ) (i. and ii. show representative images of isolated islets from control and ZnT8BKO mice; scale bar: 500 μm). b. islet number per pancreatic slice area (total pixel count). c. beta cell mass (i. and ii. show representative images of insulin staining; scale bar: 200 μm) and d. alpha cell mass (i. and ii. show representative images of glucagon staining; scale bar: 200 μm) in islets from control and ZnT8BKO mice (AU: area unit; n=2). e. Electron micrographs of isolated islets from control and ZnT8BKO mice. Manual quantifications were performed on 20 sections from 5 islets per mouse (n=2, scale bar: 500 nm).
    Figure Legend Snippet: Islet and beta cell morphological analysis in ZnT8BKO mice. a. Islet size (μm 2 ) (i. and ii. show representative images of isolated islets from control and ZnT8BKO mice; scale bar: 500 μm). b. islet number per pancreatic slice area (total pixel count). c. beta cell mass (i. and ii. show representative images of insulin staining; scale bar: 200 μm) and d. alpha cell mass (i. and ii. show representative images of glucagon staining; scale bar: 200 μm) in islets from control and ZnT8BKO mice (AU: area unit; n=2). e. Electron micrographs of isolated islets from control and ZnT8BKO mice. Manual quantifications were performed on 20 sections from 5 islets per mouse (n=2, scale bar: 500 nm).

    Techniques Used: Mouse Assay, Isolation, Staining

    ZnT8 expression in beta and alpha cells of mouse islets. a. Dispersed islet cells were immunostained for ZnT8 (red) and insulin (green) [upper panel] or glucagon (green) [lower panel]. Yellow on merged images indicates colocalization of ZnT8 and insulin or glucagon. Scale bar: 20 μm. b. Electron micrographs of immuno-gold labelled insulin [upper panel] and ZnT8 (arrows) [lower panel] in mouse islet cells. Scale bar: 400 nm [left panels]; 200 nm [right panels].
    Figure Legend Snippet: ZnT8 expression in beta and alpha cells of mouse islets. a. Dispersed islet cells were immunostained for ZnT8 (red) and insulin (green) [upper panel] or glucagon (green) [lower panel]. Yellow on merged images indicates colocalization of ZnT8 and insulin or glucagon. Scale bar: 20 μm. b. Electron micrographs of immuno-gold labelled insulin [upper panel] and ZnT8 (arrows) [lower panel] in mouse islet cells. Scale bar: 400 nm [left panels]; 200 nm [right panels].

    Techniques Used: Expressing

    11) Product Images from "FLT3 mutations in canine acute lymphocytic leukemia"

    Article Title: FLT3 mutations in canine acute lymphocytic leukemia

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-11-38

    Downstream mediators of FLT3 ITD . Protein levels of FLT3, phospho-FLT3, phospho-STAT5 and phospho-ERK were assayed by Western blotting. Protein was harvested from cell lines that were grown both with and without lestaurtinib. Beta-actin was used as a loading control.
    Figure Legend Snippet: Downstream mediators of FLT3 ITD . Protein levels of FLT3, phospho-FLT3, phospho-STAT5 and phospho-ERK were assayed by Western blotting. Protein was harvested from cell lines that were grown both with and without lestaurtinib. Beta-actin was used as a loading control.

    Techniques Used: Western Blot

    12) Product Images from "BST2/Tetherin Inhibition of Alphavirus Exit"

    Article Title: BST2/Tetherin Inhibition of Alphavirus Exit

    Journal: Viruses

    doi: 10.3390/v7042147

    Validation of Tet-On AU1-tetherin cells. ( A ) Induction of tetherin expression. Tet-On AU1-tetherin cells were incubated in the presence or absence of tetracycline (TET) for 16 h. Cell surface tetherin was detected by immunostaining of fixed, non-permeabilized cells with a tetherin-specific pAb (green) and nuclei were counterstained using Hoechst (blue). Epifluorescence microscopy images are representative of three independent experiments. Bar = 20 µm; ( B ) Ebola VLP release assay. Tet-On AU1-tetherin cells were incubated with or without tetracycline for 16 h and then transfected with an Ebola VP40-FLAG-expressing plasmid. At 48 h post-transfection, VLP were pelleted and cells were lysed, and proteins detected by SDS-PAGE and WB using anti-FLAG mAb, anti-tetherin pAb, and anti-Beta-actin mAb (loading control).
    Figure Legend Snippet: Validation of Tet-On AU1-tetherin cells. ( A ) Induction of tetherin expression. Tet-On AU1-tetherin cells were incubated in the presence or absence of tetracycline (TET) for 16 h. Cell surface tetherin was detected by immunostaining of fixed, non-permeabilized cells with a tetherin-specific pAb (green) and nuclei were counterstained using Hoechst (blue). Epifluorescence microscopy images are representative of three independent experiments. Bar = 20 µm; ( B ) Ebola VLP release assay. Tet-On AU1-tetherin cells were incubated with or without tetracycline for 16 h and then transfected with an Ebola VP40-FLAG-expressing plasmid. At 48 h post-transfection, VLP were pelleted and cells were lysed, and proteins detected by SDS-PAGE and WB using anti-FLAG mAb, anti-tetherin pAb, and anti-Beta-actin mAb (loading control).

    Techniques Used: Expressing, Incubation, Immunostaining, Epifluorescence Microscopy, Release Assay, Transfection, Plasmid Preparation, SDS Page, Western Blot

    13) Product Images from "Neuroprotection via RNA-binding protein RBM3 expression is regulated by hypothermia but not by hypoxia in human SK-N-SH neurons"

    Article Title: Neuroprotection via RNA-binding protein RBM3 expression is regulated by hypothermia but not by hypoxia in human SK-N-SH neurons

    Journal: Hypoxia

    doi: 10.2147/HP.S132462

    Western blots and densitometric quantification of RBM3 protein versus beta-actin in SK-N-SH cells after ( A ) 24 hours of hypoxia (0.2% O 2 ) or ( B ) 24 hours of subatmospheric oxygen tension (8% O 2 ) followed by 24, 48, or 72 hours of normothermia/hypothermia (total 48, 72 and 96 hours, n=4, * p
    Figure Legend Snippet: Western blots and densitometric quantification of RBM3 protein versus beta-actin in SK-N-SH cells after ( A ) 24 hours of hypoxia (0.2% O 2 ) or ( B ) 24 hours of subatmospheric oxygen tension (8% O 2 ) followed by 24, 48, or 72 hours of normothermia/hypothermia (total 48, 72 and 96 hours, n=4, * p

    Techniques Used: Western Blot

    Western blots and densitometric quantification of RBM3 protein versus beta-actin in SK-N-SH after ( A ) 48 hours of normothermia (37°C) versus hypothermia (33.5°C) and after ( B ) 24 hours of 0.2% or 8% O 2 tension compared to 24 hours of atmospheric normoxia (n=4–8, * p
    Figure Legend Snippet: Western blots and densitometric quantification of RBM3 protein versus beta-actin in SK-N-SH after ( A ) 48 hours of normothermia (37°C) versus hypothermia (33.5°C) and after ( B ) 24 hours of 0.2% or 8% O 2 tension compared to 24 hours of atmospheric normoxia (n=4–8, * p

    Techniques Used: Western Blot

    Cell morphology after 24 hours of atmospheric normoxia, subatmospheric oxygen tension, or hypoxia (21%, 8%, or 0.2% O 2 ) followed by 72 hours of normothermia (37°C) or hypothermia (33.5°C). SK-N-SH cells are stained with anti-beta III-Tubulin (red) and DAPI (blue). Abbreviation: DAPI, 4′,6-diamidin-2-phenylindol.
    Figure Legend Snippet: Cell morphology after 24 hours of atmospheric normoxia, subatmospheric oxygen tension, or hypoxia (21%, 8%, or 0.2% O 2 ) followed by 72 hours of normothermia (37°C) or hypothermia (33.5°C). SK-N-SH cells are stained with anti-beta III-Tubulin (red) and DAPI (blue). Abbreviation: DAPI, 4′,6-diamidin-2-phenylindol.

    Techniques Used: Staining

    14) Product Images from "Therapeutic Delivery of miR-148a Suppresses Ventricular Dilation in Heart Failure"

    Article Title: Therapeutic Delivery of miR-148a Suppresses Ventricular Dilation in Heart Failure

    Journal: Molecular Therapy

    doi: 10.1016/j.ymthe.2018.11.011

    miR-148a Silencing Promotes Spontaneous Eccentric Hypertrophic Remodeling and Dilated Cardiomyopathy In Vivo (A) Design of the study. Two-month-old wild-type mice were injected with a control antagomir targeting C. elegans miR-39 (antagomir -Cel-39 ) or antagomir against mmu-miR-148a-3p and subjected to sham or TAC surgery. Cardiac geometry and function was determined by serial Doppler echocardiography at 3 weeks and 6 weeks after surgery. (B) RT-PCR analysis of miR-148a-3p and unrelated let-7b expression in hearts from mice receiving control antagomir (antagomir- Cel -39) or antagomir against miR-148a . (C) Representative images of whole hearts (top panel), H E-stained sections of four-chamber view (second panel), high-magnification H E-stained sections (third panel), Sirius-red-stained sections (fourth panel), and WGA-stained (fifth panel) histological sections. (D) Measurement of LV mass/BW ratio, (E) LVPWs, (F) LVIDs, or (G) EF by echocardiography in antagomir control and antagomir-148a-treated mice after sham or TAC surgery. Measurements of (H) fibrotic area (Sirius red staining) and (I) cell-surface area in control and antagomir-148a-treated mice after sham or TAC surgery. (J) RT-PCR analysis of transcript abundance of the hypertrophic stress marker genes Nppa , Nppb , Acta1 , and Myh7 in hearts from control antagomir or antagomir-148a-treated mice after sham or TAC surgery. (K–M) Western blot analysis (K) and quantification of myocardial gp130 (L) or phosphorylated and unphosphorylated forms of STAT3 (M) in hearts from mice subjected to sham or TAC surgery and receiving antagomir-148a or a control antagomir (antagomir- Cel -39). Data are means ± SEM. One-way ANOVA with Bonferroni’s or Newman-Keuls multiple-comparison test was used to compare groups. n, number of hearts; TAC, transverse aortic constriction; WGA, wheat germ agglutinin; LV mass/BW, left ventricular mass to body weight ratio; LVPWs, left ventricular posterior wall end systole; LVIDs, left ventricular internal diameter in systole; EF, ejection fraction; Nppa , natriuretic peptide type A; Nppb , natriuretic peptide type B; Acta1 , alpha skeletal actin; Myh7 , beta myosin heavy chain. *p
    Figure Legend Snippet: miR-148a Silencing Promotes Spontaneous Eccentric Hypertrophic Remodeling and Dilated Cardiomyopathy In Vivo (A) Design of the study. Two-month-old wild-type mice were injected with a control antagomir targeting C. elegans miR-39 (antagomir -Cel-39 ) or antagomir against mmu-miR-148a-3p and subjected to sham or TAC surgery. Cardiac geometry and function was determined by serial Doppler echocardiography at 3 weeks and 6 weeks after surgery. (B) RT-PCR analysis of miR-148a-3p and unrelated let-7b expression in hearts from mice receiving control antagomir (antagomir- Cel -39) or antagomir against miR-148a . (C) Representative images of whole hearts (top panel), H E-stained sections of four-chamber view (second panel), high-magnification H E-stained sections (third panel), Sirius-red-stained sections (fourth panel), and WGA-stained (fifth panel) histological sections. (D) Measurement of LV mass/BW ratio, (E) LVPWs, (F) LVIDs, or (G) EF by echocardiography in antagomir control and antagomir-148a-treated mice after sham or TAC surgery. Measurements of (H) fibrotic area (Sirius red staining) and (I) cell-surface area in control and antagomir-148a-treated mice after sham or TAC surgery. (J) RT-PCR analysis of transcript abundance of the hypertrophic stress marker genes Nppa , Nppb , Acta1 , and Myh7 in hearts from control antagomir or antagomir-148a-treated mice after sham or TAC surgery. (K–M) Western blot analysis (K) and quantification of myocardial gp130 (L) or phosphorylated and unphosphorylated forms of STAT3 (M) in hearts from mice subjected to sham or TAC surgery and receiving antagomir-148a or a control antagomir (antagomir- Cel -39). Data are means ± SEM. One-way ANOVA with Bonferroni’s or Newman-Keuls multiple-comparison test was used to compare groups. n, number of hearts; TAC, transverse aortic constriction; WGA, wheat germ agglutinin; LV mass/BW, left ventricular mass to body weight ratio; LVPWs, left ventricular posterior wall end systole; LVIDs, left ventricular internal diameter in systole; EF, ejection fraction; Nppa , natriuretic peptide type A; Nppb , natriuretic peptide type B; Acta1 , alpha skeletal actin; Myh7 , beta myosin heavy chain. *p

    Techniques Used: In Vivo, Mouse Assay, Injection, Reverse Transcription Polymerase Chain Reaction, Expressing, Staining, Whole Genome Amplification, Marker, Western Blot

    Cardiac AAV9-miR-148a Overexpression Protects the Heart against Eccentric Hypertrophy, Chamber Dilation, and Systolic Dysfunction (A) Design of the study. Two-month-old mice were injected with control AAV9 (AAV9-MCS) or an AAV9 designed to overexpress mmu-miR-148a-3p (AAV9-148a) and subjected to sham or TAC surgery. Cardiac geometry and function was determined by serial Doppler echocardiography at 3 and 6 weeks after surgery. (B) RT-PCR analysis of miR-148a-3p expression in hearts from mice receiving AAV9-MCS or AAV9-148a virus. (C) Representative images of whole hearts (top panel), H E-stained sections of four-chamber view (second panel), high-magnification H E-stained sections (third panel), Sirius-red-stained sections (fourth panel), and WGA-stained (fifth panel) histological sections. (D) Measurements of LV mass/BW ratio, (E) LVPWs, (F) LVIDs, (G) EF, (H) fibrotic area (Sirius red staining), and (I) cell-surface area in AAV9-CMV- and AAV9-148a-treated mice after sham or TAC surgery. (J) RT-PCR analysis of transcript abundance of hypertrophic stress marker genes Nppa , Nppb , Acta1 , and Myh7 in hearts from AAV9-MCS and AAV9-148a-treated mice after sham or TAC surgery. (K–M) Western blot analysis (K) and quantification of myocardial gp130 (L) or phosphorylated and unphosphorylated forms of STAT3 (M) in hearts from mice subjected to sham or TAC surgery and receiving AAV9-148a or AAV9-MCS (MCS, multiple cloning site; empty vector). Data are means ± SEM. One-way ANOVA with Bonferroni’s or Newman-Keul’s multiple-comparison test was used to compare groups. n, number of hearts; AAV9, adeno-associated vector serotype 9; TAC, transverse aortic constriction; WGA, wheat germ agglutinin; LV mass/BW, left ventricular mass to body weight ratio; LVPWs, left ventricular posterior wall end systole; LVIDs, left ventricular internal diameter in systole; EF, ejection fraction; Nppa , natriuretic peptide type A; Nppb , natriuretic peptide type B; Acta1 , alpha skeletal actin; Myh7 , beta myosin heavy chain. *p
    Figure Legend Snippet: Cardiac AAV9-miR-148a Overexpression Protects the Heart against Eccentric Hypertrophy, Chamber Dilation, and Systolic Dysfunction (A) Design of the study. Two-month-old mice were injected with control AAV9 (AAV9-MCS) or an AAV9 designed to overexpress mmu-miR-148a-3p (AAV9-148a) and subjected to sham or TAC surgery. Cardiac geometry and function was determined by serial Doppler echocardiography at 3 and 6 weeks after surgery. (B) RT-PCR analysis of miR-148a-3p expression in hearts from mice receiving AAV9-MCS or AAV9-148a virus. (C) Representative images of whole hearts (top panel), H E-stained sections of four-chamber view (second panel), high-magnification H E-stained sections (third panel), Sirius-red-stained sections (fourth panel), and WGA-stained (fifth panel) histological sections. (D) Measurements of LV mass/BW ratio, (E) LVPWs, (F) LVIDs, (G) EF, (H) fibrotic area (Sirius red staining), and (I) cell-surface area in AAV9-CMV- and AAV9-148a-treated mice after sham or TAC surgery. (J) RT-PCR analysis of transcript abundance of hypertrophic stress marker genes Nppa , Nppb , Acta1 , and Myh7 in hearts from AAV9-MCS and AAV9-148a-treated mice after sham or TAC surgery. (K–M) Western blot analysis (K) and quantification of myocardial gp130 (L) or phosphorylated and unphosphorylated forms of STAT3 (M) in hearts from mice subjected to sham or TAC surgery and receiving AAV9-148a or AAV9-MCS (MCS, multiple cloning site; empty vector). Data are means ± SEM. One-way ANOVA with Bonferroni’s or Newman-Keul’s multiple-comparison test was used to compare groups. n, number of hearts; AAV9, adeno-associated vector serotype 9; TAC, transverse aortic constriction; WGA, wheat germ agglutinin; LV mass/BW, left ventricular mass to body weight ratio; LVPWs, left ventricular posterior wall end systole; LVIDs, left ventricular internal diameter in systole; EF, ejection fraction; Nppa , natriuretic peptide type A; Nppb , natriuretic peptide type B; Acta1 , alpha skeletal actin; Myh7 , beta myosin heavy chain. *p

    Techniques Used: Over Expression, Mouse Assay, Injection, Reverse Transcription Polymerase Chain Reaction, Expressing, Staining, Whole Genome Amplification, Marker, Western Blot, Clone Assay, Plasmid Preparation

    miR-148a Overexpression Prevents the Transition of Pressure-Overload-Induced Concentric Hypertrophic Remodeling toward Eccentric Hypertrophy (A) Design of the study. Two-month-old mice were subjected to sham or TAC surgery and 4 weeks later injected with control AAV9 (AAV9-CMV) or AAV9-148a. Cardiac geometry and function was determined by serial Doppler echocardiography at 3, 5, and 7 weeks after surgery. (B) RT-PCR analysis of miR-148a-3p expression in hearts from mice receiving AAV9-CMV or AAV9-148a virus. (C) Representative images of H E-stained sections of four-chamber view (top panel), high-magnification H E-stained sections (second panel), Sirius-red-stained sections (third panel), and WGA-stained (fourth panel) histological sections. (D) Measurements of LV mass/BW ratio, (E) LVPWs, (F) LVIDs, (G) EF, (H) fibrotic area (Sirius red staining), and (I) cell surface area in AAV9-CMV and AAV9-148a-treated mice after sham or TAC surgery. (J) RT-PCR analysis of transcript abundance of hypertrophic stress marker genes Nppa , Nppb , Acta1 , and Myh7 in hearts from AAV9-CMV and AAV9-148a-treated mice after sham or TAC surgery. (K) Model depicting the influence of miR-148a on the CT-1-gp130-STAT3 signaling axis in cardiac stress situations, resulting in differential activation of STAT3 signaling and either concentric or eccentric remodeling. Data are means ± SEM. One-way ANOVA with Bonferroni’s multiple-comparison test was used to compare groups. n, number of hearts; AAV9, adeno-associated vector serotype 9; TAC, transverse aortic constriction; WGA, wheat germ agglutinin; LV mass/BW, left ventricular mass to body weight ratio; LVPWs, left ventricular posterior wall end systole; LVIDs, left ventricular internal diameter in systole; EF, ejection fraction; Nppa , natriuretic peptide type A; Nppb , natriuretic peptide type B; Acta1 , alpha skeletal actin; Myh7 , beta myosin heavy chain. *p
    Figure Legend Snippet: miR-148a Overexpression Prevents the Transition of Pressure-Overload-Induced Concentric Hypertrophic Remodeling toward Eccentric Hypertrophy (A) Design of the study. Two-month-old mice were subjected to sham or TAC surgery and 4 weeks later injected with control AAV9 (AAV9-CMV) or AAV9-148a. Cardiac geometry and function was determined by serial Doppler echocardiography at 3, 5, and 7 weeks after surgery. (B) RT-PCR analysis of miR-148a-3p expression in hearts from mice receiving AAV9-CMV or AAV9-148a virus. (C) Representative images of H E-stained sections of four-chamber view (top panel), high-magnification H E-stained sections (second panel), Sirius-red-stained sections (third panel), and WGA-stained (fourth panel) histological sections. (D) Measurements of LV mass/BW ratio, (E) LVPWs, (F) LVIDs, (G) EF, (H) fibrotic area (Sirius red staining), and (I) cell surface area in AAV9-CMV and AAV9-148a-treated mice after sham or TAC surgery. (J) RT-PCR analysis of transcript abundance of hypertrophic stress marker genes Nppa , Nppb , Acta1 , and Myh7 in hearts from AAV9-CMV and AAV9-148a-treated mice after sham or TAC surgery. (K) Model depicting the influence of miR-148a on the CT-1-gp130-STAT3 signaling axis in cardiac stress situations, resulting in differential activation of STAT3 signaling and either concentric or eccentric remodeling. Data are means ± SEM. One-way ANOVA with Bonferroni’s multiple-comparison test was used to compare groups. n, number of hearts; AAV9, adeno-associated vector serotype 9; TAC, transverse aortic constriction; WGA, wheat germ agglutinin; LV mass/BW, left ventricular mass to body weight ratio; LVPWs, left ventricular posterior wall end systole; LVIDs, left ventricular internal diameter in systole; EF, ejection fraction; Nppa , natriuretic peptide type A; Nppb , natriuretic peptide type B; Acta1 , alpha skeletal actin; Myh7 , beta myosin heavy chain. *p

    Techniques Used: Over Expression, Mouse Assay, Injection, Reverse Transcription Polymerase Chain Reaction, Expressing, Staining, Whole Genome Amplification, Marker, Activation Assay, Plasmid Preparation

    15) Product Images from "Difference in the Cytotoxic Effects of Toxin B from Clostridium difficile Strain VPI 10463 and Toxin B from Variant Clostridium difficile Strain 1470 ▿"

    Article Title: Difference in the Cytotoxic Effects of Toxin B from Clostridium difficile Strain VPI 10463 and Toxin B from Variant Clostridium difficile Strain 1470 ▿

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01705-06

    Cytotoxic effect in nonsynchronized RBL cells. (A) Activation of caspase-3 by TcdB. RBL cells were exposed to increasing concentrations of TcdB (▪) or TcdBF (•) for 3 h. The cellular levels of procaspase-3 and caspase-3 were determined by Western blot analysis. The signal intensity was normalized to the intensity of the beta-actin signal. (B) Cytotoxic effect of TcdB. RBL cells were exposed to increasing concentrations of TcdB for 24 h. The cytopathic effect was determined as the percent rounded cells. Annexin V-positive cells were stained with annexin V-Alexa Fluor 488 and visualized by fluorescence microscopy. (C) Cytotoxic effect of TcdB. RBL cells were exposed to increasing concentrations of TcdBF for 24 h. The cytopathic effect was determined as the percent rounded cells. Annexin V-positive cells were stained with annexin V-Alexa Fluor 488 and visualized by fluorescence microscopy. (D) Cytotoxic effect in nonsynchronized RBL cells. RBL cells were exposed to latrunculin B (2.5 μM), staurosporine (STS) (0.2 μM), TcdB (0.1 μg/ml), or TcdBF (1 μg/ml) or left untreated for 24 h. Phosphatidylserine exposure was visualized by annexin V staining.
    Figure Legend Snippet: Cytotoxic effect in nonsynchronized RBL cells. (A) Activation of caspase-3 by TcdB. RBL cells were exposed to increasing concentrations of TcdB (▪) or TcdBF (•) for 3 h. The cellular levels of procaspase-3 and caspase-3 were determined by Western blot analysis. The signal intensity was normalized to the intensity of the beta-actin signal. (B) Cytotoxic effect of TcdB. RBL cells were exposed to increasing concentrations of TcdB for 24 h. The cytopathic effect was determined as the percent rounded cells. Annexin V-positive cells were stained with annexin V-Alexa Fluor 488 and visualized by fluorescence microscopy. (C) Cytotoxic effect of TcdB. RBL cells were exposed to increasing concentrations of TcdBF for 24 h. The cytopathic effect was determined as the percent rounded cells. Annexin V-positive cells were stained with annexin V-Alexa Fluor 488 and visualized by fluorescence microscopy. (D) Cytotoxic effect in nonsynchronized RBL cells. RBL cells were exposed to latrunculin B (2.5 μM), staurosporine (STS) (0.2 μM), TcdB (0.1 μg/ml), or TcdBF (1 μg/ml) or left untreated for 24 h. Phosphatidylserine exposure was visualized by annexin V staining.

    Techniques Used: Activation Assay, Western Blot, Staining, Fluorescence, Microscopy

    Cellular activity of TcdB and TcdBF in RBL cells. (A) Cytopathic effect of TcdB and TcdBF. RBL cells were exposed to TcdB (▪) or TcdBF (•) for 3 h. The cytopathic effect was determined as percent rounded cells. (B to E) Influence of the toxins on the cellular levels of Rho proteins. RBL cells were exposed to TcdB (▪) or TcdBF (•) for 3 h. The cellular levels of Rho proteins were determined by Western blotting with the indicated antibodies. The signal intensity was normalized to the intensity of the corresponding beta-actin signal.
    Figure Legend Snippet: Cellular activity of TcdB and TcdBF in RBL cells. (A) Cytopathic effect of TcdB and TcdBF. RBL cells were exposed to TcdB (▪) or TcdBF (•) for 3 h. The cytopathic effect was determined as percent rounded cells. (B to E) Influence of the toxins on the cellular levels of Rho proteins. RBL cells were exposed to TcdB (▪) or TcdBF (•) for 3 h. The cellular levels of Rho proteins were determined by Western blotting with the indicated antibodies. The signal intensity was normalized to the intensity of the corresponding beta-actin signal.

    Techniques Used: Activity Assay, Western Blot

    16) Product Images from "The MAPK Pathway Signals Telomerase Modulation in Response to Isothiocyanate-Induced DNA Damage of Human Liver Cancer Cells"

    Article Title: The MAPK Pathway Signals Telomerase Modulation in Response to Isothiocyanate-Induced DNA Damage of Human Liver Cancer Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0053240

    Apoptosis induction and telomerase loss in HepG2 cells are independent from ROS production. a) Effect of MTBITC on ROS production in adherent growing cells. Cells were exposed to MTBITC for 1 to 6 h, washed thoroughly with PBS and subsequently exposed to 100 µM spin probe CMH in Krebs-Hepes buffer. ROS were then quantified using ESR spectrometry. Positive control: 100 µM menadione. The pictures display the middle peak of ESR spectra of CMH spin probe labelled samples. Bars are mean ± SD, n = 3. b) Representative immunoblot for HNE Lys-adducts after exposure to MTBITC or DMSO for different time points. Total lysate of HepG2 cells was subjected to immunoblotting using an anti-HNE Lys antibody. Exposure of cells to HNE for 80 min. was used as positive control. The blot was reprobed with anti-β-actin antibody to ensure equal protein loading. c+d) Effect of NAC pre-treatment. HepG2 cells were pre-treated with NAC for 1 h, washed with PBS and subsequently exposed to 25 µM MTBITC or 0.1% DMSO for another 24 h. Cells were then prepared for (c) TRAP analysis or (d) flow cytometric analysis of the mitochondrial membrane potential (MMP) as parameter for apoptosis; *p≤0.05. M: 50 bp DNA marker; 1) 0.1% DMSO 2) 25 µM MTBITC 3) 5 mM NAC +0.1% DMSO,4) 5 mM NAC +25 µM MTBITC 5) cell lytic buffer 6) destilled water 7) 0.1% DMSO, heat treated. e) Effect of 25 µM MTBITC on mtDNA damage after 1 or 24 h exposure of HepG2 cells. Agarose gel electrophoresis of amplified mtDNA multiplex PCR products is shown in representative image details. Each lane contains amplification products obtained with mtDNA from one single cell using specific PCR primers. As positive control for mtDNA damage, cells were exposed to 10 min UV irradiation. f ) Representative pictures of beta-galactosidase staining of HCC cells after exposure to MTBITC for 72 h, captured by light microscopy. beta-galactosidase positive cells are reflected by dark color of the cells. SC: solvent control = 0.1% DMSO. Scale bar = 200 µm, all panels have the same magnification.
    Figure Legend Snippet: Apoptosis induction and telomerase loss in HepG2 cells are independent from ROS production. a) Effect of MTBITC on ROS production in adherent growing cells. Cells were exposed to MTBITC for 1 to 6 h, washed thoroughly with PBS and subsequently exposed to 100 µM spin probe CMH in Krebs-Hepes buffer. ROS were then quantified using ESR spectrometry. Positive control: 100 µM menadione. The pictures display the middle peak of ESR spectra of CMH spin probe labelled samples. Bars are mean ± SD, n = 3. b) Representative immunoblot for HNE Lys-adducts after exposure to MTBITC or DMSO for different time points. Total lysate of HepG2 cells was subjected to immunoblotting using an anti-HNE Lys antibody. Exposure of cells to HNE for 80 min. was used as positive control. The blot was reprobed with anti-β-actin antibody to ensure equal protein loading. c+d) Effect of NAC pre-treatment. HepG2 cells were pre-treated with NAC for 1 h, washed with PBS and subsequently exposed to 25 µM MTBITC or 0.1% DMSO for another 24 h. Cells were then prepared for (c) TRAP analysis or (d) flow cytometric analysis of the mitochondrial membrane potential (MMP) as parameter for apoptosis; *p≤0.05. M: 50 bp DNA marker; 1) 0.1% DMSO 2) 25 µM MTBITC 3) 5 mM NAC +0.1% DMSO,4) 5 mM NAC +25 µM MTBITC 5) cell lytic buffer 6) destilled water 7) 0.1% DMSO, heat treated. e) Effect of 25 µM MTBITC on mtDNA damage after 1 or 24 h exposure of HepG2 cells. Agarose gel electrophoresis of amplified mtDNA multiplex PCR products is shown in representative image details. Each lane contains amplification products obtained with mtDNA from one single cell using specific PCR primers. As positive control for mtDNA damage, cells were exposed to 10 min UV irradiation. f ) Representative pictures of beta-galactosidase staining of HCC cells after exposure to MTBITC for 72 h, captured by light microscopy. beta-galactosidase positive cells are reflected by dark color of the cells. SC: solvent control = 0.1% DMSO. Scale bar = 200 µm, all panels have the same magnification.

    Techniques Used: Electron Paramagnetic Resonance, Positive Control, Flow Cytometry, Marker, Agarose Gel Electrophoresis, Amplification, Multiplex Assay, Polymerase Chain Reaction, Irradiation, Staining, Light Microscopy

    17) Product Images from "A Novel Human Pluripotent Stem Cell-Derived Neural Crest Model of Treacher Collins Syndrome Shows Defects in Cell Death and Migration"

    Article Title: A Novel Human Pluripotent Stem Cell-Derived Neural Crest Model of Treacher Collins Syndrome Shows Defects in Cell Death and Migration

    Journal: Stem Cells and Development

    doi: 10.1089/scd.2017.0234

    Differentiation of HESC-derived NC to their derivatives. (A) HESC-derived NC cells were plated on polyornithine-laminin-coated culture dishes to induce neuronal differentiation. (A.i, A.ii) Beta III Tubulin immunocytochemistry showing differentiation to peripheral neurons. (A.iii) NC cells stained with beta III Tubulin as a negative control. (A.iv) Magnification of the white square in (A.ii) . White scale bar: 100 μm, blue scale bar: 50 μm. Representative images of three independent biological replicates are shown. (B) Immunocytochemistry of the SMC proteins CNN1, ACTA2, and SM22A following HESC-derived NC differentiation to SMC with PDGF-BB (10 ng/mL) and TGF-β1 (2 ng/mL) for 12 days (12D PT). White scale bar: 100 μm, red scale bar: 20 μm. Representative images of three independent biological replicates. (C) qRT-PCR showing the upregulation of the specific melanocyte genes MITF and KIT and the downregulation of specific NC genes FOXD3 and P75 in NC P7 derived melanocytes (NC-MEL). The relative mRNA level was normalized to the housekeeping gene PBGD . Results are presented as mean ± SD of three independent experiments. * P
    Figure Legend Snippet: Differentiation of HESC-derived NC to their derivatives. (A) HESC-derived NC cells were plated on polyornithine-laminin-coated culture dishes to induce neuronal differentiation. (A.i, A.ii) Beta III Tubulin immunocytochemistry showing differentiation to peripheral neurons. (A.iii) NC cells stained with beta III Tubulin as a negative control. (A.iv) Magnification of the white square in (A.ii) . White scale bar: 100 μm, blue scale bar: 50 μm. Representative images of three independent biological replicates are shown. (B) Immunocytochemistry of the SMC proteins CNN1, ACTA2, and SM22A following HESC-derived NC differentiation to SMC with PDGF-BB (10 ng/mL) and TGF-β1 (2 ng/mL) for 12 days (12D PT). White scale bar: 100 μm, red scale bar: 20 μm. Representative images of three independent biological replicates. (C) qRT-PCR showing the upregulation of the specific melanocyte genes MITF and KIT and the downregulation of specific NC genes FOXD3 and P75 in NC P7 derived melanocytes (NC-MEL). The relative mRNA level was normalized to the housekeeping gene PBGD . Results are presented as mean ± SD of three independent experiments. * P

    Techniques Used: Derivative Assay, Immunocytochemistry, Staining, Negative Control, Quantitative RT-PCR

    18) Product Images from "Cortactin Is a Substrate of Activated Cdc42-Associated Kinase 1 (ACK1) during Ligand-induced Epidermal Growth Factor Receptor Downregulation"

    Article Title: Cortactin Is a Substrate of Activated Cdc42-Associated Kinase 1 (ACK1) during Ligand-induced Epidermal Growth Factor Receptor Downregulation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0044363

    ACK1 directly phosphorylates cortactin at Y421/466/482. (A) Characterization of purified ACK1. Recombinant 6X-histidine tagged recombinant ACK1 was purified from SF9 cells and 5 micrograms stained with either Coomassie blue ( left ) or analyzed by Western blotting with anti-ACK antibodies ( middle ). Kinase activity was assayed by analyzing autophosphorylation (Autophos) using 0.5 micrograms of 6X-His ACK1 incubated with gamma- 32 P-ATP for 10 min at 30°C. The reaction was evaluated by SDS-PAGE and autoradiography ( right ). The position of molecular weight standards is shown on the left and 6X-His ACK1 on the right. (B) Identification of cortactin tyrosine residues phosphorylated by ACK1. Purified GST-cortactin murine wild type (WT) or Y421/466/482F (triple tyrosine mutant; TYM) (0.5 micrograms) were incubated in kinase assays without or with 3U of purified Src ( left ) or with the indicated amounts of purified 6X-His ACK1 ( middle and right ) in the presence of [gamma- 32 P]ATP. Reactions were separated by SDS-PAGE and analyzed by audioradiography. Arrows indicate the positions of phosphorylated GST-cortactin and 6X-His ACK1 ( left ). (C) Determination of the K M of ACK1 for cortactin. Graphical representation showing ACK1 phosphorylation kinetics for cortactin calculated from the kinase assay in B ( right panel). Percent of maximum phosphorylation signal measured by densitometry is represented on the ordinate versus concentration of ACK1 (in micrograms) on the abscissa. The calculated K M for cortactin phosphorylation is shown. Data in B and C are representative from three independent experiments. (D) ACK1 knockdown reduces cortactin phosphorylation on tyrosine 421. 1483 cells transfected with scrambled control (Ctl) siRNA or ACK1-targeting siRNA were lysed and analyzed by Western blotting for ACK1 knockdown (ACK1) and cortactin pY421 phosphorylation (Cort pY421). Beta actin was blotted to verify equivalent protein loading. Blots are representative of two independent experiments.
    Figure Legend Snippet: ACK1 directly phosphorylates cortactin at Y421/466/482. (A) Characterization of purified ACK1. Recombinant 6X-histidine tagged recombinant ACK1 was purified from SF9 cells and 5 micrograms stained with either Coomassie blue ( left ) or analyzed by Western blotting with anti-ACK antibodies ( middle ). Kinase activity was assayed by analyzing autophosphorylation (Autophos) using 0.5 micrograms of 6X-His ACK1 incubated with gamma- 32 P-ATP for 10 min at 30°C. The reaction was evaluated by SDS-PAGE and autoradiography ( right ). The position of molecular weight standards is shown on the left and 6X-His ACK1 on the right. (B) Identification of cortactin tyrosine residues phosphorylated by ACK1. Purified GST-cortactin murine wild type (WT) or Y421/466/482F (triple tyrosine mutant; TYM) (0.5 micrograms) were incubated in kinase assays without or with 3U of purified Src ( left ) or with the indicated amounts of purified 6X-His ACK1 ( middle and right ) in the presence of [gamma- 32 P]ATP. Reactions were separated by SDS-PAGE and analyzed by audioradiography. Arrows indicate the positions of phosphorylated GST-cortactin and 6X-His ACK1 ( left ). (C) Determination of the K M of ACK1 for cortactin. Graphical representation showing ACK1 phosphorylation kinetics for cortactin calculated from the kinase assay in B ( right panel). Percent of maximum phosphorylation signal measured by densitometry is represented on the ordinate versus concentration of ACK1 (in micrograms) on the abscissa. The calculated K M for cortactin phosphorylation is shown. Data in B and C are representative from three independent experiments. (D) ACK1 knockdown reduces cortactin phosphorylation on tyrosine 421. 1483 cells transfected with scrambled control (Ctl) siRNA or ACK1-targeting siRNA were lysed and analyzed by Western blotting for ACK1 knockdown (ACK1) and cortactin pY421 phosphorylation (Cort pY421). Beta actin was blotted to verify equivalent protein loading. Blots are representative of two independent experiments.

    Techniques Used: Purification, Recombinant, Staining, Western Blot, Activity Assay, Incubation, SDS Page, Autoradiography, Molecular Weight, Mutagenesis, Kinase Assay, Concentration Assay, Transfection, CTL Assay

    19) Product Images from "STAT2 Mediates Innate Immunity to Dengue Virus in the Absence of STAT1 via the Type I Interferon Receptor"

    Article Title: STAT2 Mediates Innate Immunity to Dengue Virus in the Absence of STAT1 via the Type I Interferon Receptor

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1001297

    Activation of STAT1 and STAT2 in bone marrow-derived macrophages. (A–C) Whole cell lysates were generated from bone marrow-derived macrophages that were isolated from WT, STAT1 −/− , STAT1 −/− /2 −/− , or STAT1 −/− /AR −/− mice and treated with (A) 1000U/mL recombinant murine IFN-β for 15 min, (B) serum from naïve (N) or S221-infected (I) mice for 1 hour, or (C) infected with S221 (MOI = 5) for the times indicated. Protein levels of STAT1, phosphorylated STAT1, STAT2, phosphorylated STAT2, and beta-actin were examined by Western blot. (D) Nuclear localization of STAT2 in bone marrow-derived macrophages isolated from WT and STAT1 −/− mice, and treated with serum from naïve (N) or S221-infected (I) mice for 1 hour. STAT2 (red), and DAPI (blue). Original magnification, ×400. Images are representative of each strain. Data are representative of two or more individual experiments.
    Figure Legend Snippet: Activation of STAT1 and STAT2 in bone marrow-derived macrophages. (A–C) Whole cell lysates were generated from bone marrow-derived macrophages that were isolated from WT, STAT1 −/− , STAT1 −/− /2 −/− , or STAT1 −/− /AR −/− mice and treated with (A) 1000U/mL recombinant murine IFN-β for 15 min, (B) serum from naïve (N) or S221-infected (I) mice for 1 hour, or (C) infected with S221 (MOI = 5) for the times indicated. Protein levels of STAT1, phosphorylated STAT1, STAT2, phosphorylated STAT2, and beta-actin were examined by Western blot. (D) Nuclear localization of STAT2 in bone marrow-derived macrophages isolated from WT and STAT1 −/− mice, and treated with serum from naïve (N) or S221-infected (I) mice for 1 hour. STAT2 (red), and DAPI (blue). Original magnification, ×400. Images are representative of each strain. Data are representative of two or more individual experiments.

    Techniques Used: Activation Assay, Derivative Assay, Generated, Isolation, Mouse Assay, Recombinant, Infection, Western Blot

    20) Product Images from "Inflammatory and cytotoxic effects of bifenthrin in primary microglia and organotypic hippocampal slice cultures"

    Article Title: Inflammatory and cytotoxic effects of bifenthrin in primary microglia and organotypic hippocampal slice cultures

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-018-1198-1

    BF exposure increases the expression of pro-inflammatory markers in primary microglia cells. Microglial cells were exposed to different concentrations of BF (1–20 μM) for 24 h. Gene expression of TNF-alpha ( a ), IL-6 ( b ), COX-2 ( c ), and mPGES-1 ( d ) was analyzed by real-time quantitative PCR. GAPDH was used as an internal control for normalization, and data were quantified by using the comparative cycle threshold Ct method. Similarly, cells were treated with BF and thereafter incubated with or without LPS (100 ng/mL) as a positive control (black column) for 24 h. Whole cell lysates were subjected to Western blot for COX-2 ( e , f ), mPGES-1 ( g , h ), and beta-actin. To confirm equal sample loading, beta-actin was used for normalization. Data are presented as percentage control of DMSO. Statistical analyses were carried out by using one-way ANOVA followed by post hoc Student–Newman–Keuls test. Results are expressed as means ± SEM of three independent experiments. *p
    Figure Legend Snippet: BF exposure increases the expression of pro-inflammatory markers in primary microglia cells. Microglial cells were exposed to different concentrations of BF (1–20 μM) for 24 h. Gene expression of TNF-alpha ( a ), IL-6 ( b ), COX-2 ( c ), and mPGES-1 ( d ) was analyzed by real-time quantitative PCR. GAPDH was used as an internal control for normalization, and data were quantified by using the comparative cycle threshold Ct method. Similarly, cells were treated with BF and thereafter incubated with or without LPS (100 ng/mL) as a positive control (black column) for 24 h. Whole cell lysates were subjected to Western blot for COX-2 ( e , f ), mPGES-1 ( g , h ), and beta-actin. To confirm equal sample loading, beta-actin was used for normalization. Data are presented as percentage control of DMSO. Statistical analyses were carried out by using one-way ANOVA followed by post hoc Student–Newman–Keuls test. Results are expressed as means ± SEM of three independent experiments. *p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Incubation, Positive Control, Western Blot

    BF exposure increases the expression of Nrf-2 and NF-kappaB in primary microglia. Microglial cells were exposed to different concentrations of BF (1–20 μM) for 24 h. Gene expression of Nrf-2 ( a ) and NF-kappaBp65 ( b ) was analyzed by real-time quantitative PCR. GAPDH was used as an internal control for normalization, and data were quantified by using the comparative cycle threshold Ct method. Similarly, cells were treated with BF and thereafter incubated with or without LPS (100 ng/mL) as a positive control (black column) for 24 h. Whole cell lysates were subjected to Western blot for Nrf-2 ( c , d ), NF-kappaBp65 ( e , f ), and beta-actin. Data are presented as percentage control of DMSO. Statistical analyses were carried out by using one-way ANOVA followed by post hoc Student–Newman–Keuls test. Results are expressed as means ± SEM of three independent experiments. * p
    Figure Legend Snippet: BF exposure increases the expression of Nrf-2 and NF-kappaB in primary microglia. Microglial cells were exposed to different concentrations of BF (1–20 μM) for 24 h. Gene expression of Nrf-2 ( a ) and NF-kappaBp65 ( b ) was analyzed by real-time quantitative PCR. GAPDH was used as an internal control for normalization, and data were quantified by using the comparative cycle threshold Ct method. Similarly, cells were treated with BF and thereafter incubated with or without LPS (100 ng/mL) as a positive control (black column) for 24 h. Whole cell lysates were subjected to Western blot for Nrf-2 ( c , d ), NF-kappaBp65 ( e , f ), and beta-actin. Data are presented as percentage control of DMSO. Statistical analyses were carried out by using one-way ANOVA followed by post hoc Student–Newman–Keuls test. Results are expressed as means ± SEM of three independent experiments. * p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Incubation, Positive Control, Western Blot

    21) Product Images from "The essential role of GATA transcription factors in adult murine prostate"

    Article Title: The essential role of GATA transcription factors in adult murine prostate

    Journal: Oncotarget

    doi: 10.18632/oncotarget.10294

    Expression of the GATA and other key transcription factors in WT murine prostate ( A ) Co-immunofluorescence staining of GATA2 (green), GATA3 (green) and p63 (red) in prostatic lobes. The scale bars, 20 μm in all panels. ( B ) mRNA levels of GATA2 and GATA3 in three prostatic lobes as determined by real-time qPCR. Data are average ± SE ( n = 3). ( C ) Protein levels of GATA2, GATA3, AR, and FoxA1 in prostatic lobes as determined by Western blot analysis. Because Western samples for these four transcription factors were from the same batch of prostate tissue lysates, one beta-Actin Western result was shown here to indicate an equal sample loading.
    Figure Legend Snippet: Expression of the GATA and other key transcription factors in WT murine prostate ( A ) Co-immunofluorescence staining of GATA2 (green), GATA3 (green) and p63 (red) in prostatic lobes. The scale bars, 20 μm in all panels. ( B ) mRNA levels of GATA2 and GATA3 in three prostatic lobes as determined by real-time qPCR. Data are average ± SE ( n = 3). ( C ) Protein levels of GATA2, GATA3, AR, and FoxA1 in prostatic lobes as determined by Western blot analysis. Because Western samples for these four transcription factors were from the same batch of prostate tissue lysates, one beta-Actin Western result was shown here to indicate an equal sample loading.

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

    22) Product Images from "The novel HSP90 inhibitor AT13387 potentiates radiation effects in squamous cell carcinoma and adenocarcinoma cells"

    Article Title: The novel HSP90 inhibitor AT13387 potentiates radiation effects in squamous cell carcinoma and adenocarcinoma cells

    Journal: Oncotarget

    doi:

    Effect of AT13387 treatment on HSP90 client protein levels (HSP90, ATM, DNA-PKcs, EGFR, AKT, CD44, CD44v6) A. Dose-dependent downregulation of client proteins. H314 cells and LS1474T cells treated with the indicated doses of AT13387 for 24 h. B. Recurrence of protein level after AT13387 treatment. H314 and LS174T cells were treated with 200 nM AT13387 for 24 h. After drug treatment cells were kept in drug-free complete medium for 0, 4, 8 and 24 h. Lysates were harvested and equivalent amounts of protein from each lysate were resolved by SDS-PAGE and immunoblotting with the indicated antibodies. The expression levels of beta actin were used to ensure equal loading. Above: Representative Western blots, Below: Western blot quantification. Protein levels were normalized to beta actin, and were normalized to the level of untreated control (dashed line). One way-ANOVA with Bonferroni post-test was used to calculate statistics: * p
    Figure Legend Snippet: Effect of AT13387 treatment on HSP90 client protein levels (HSP90, ATM, DNA-PKcs, EGFR, AKT, CD44, CD44v6) A. Dose-dependent downregulation of client proteins. H314 cells and LS1474T cells treated with the indicated doses of AT13387 for 24 h. B. Recurrence of protein level after AT13387 treatment. H314 and LS174T cells were treated with 200 nM AT13387 for 24 h. After drug treatment cells were kept in drug-free complete medium for 0, 4, 8 and 24 h. Lysates were harvested and equivalent amounts of protein from each lysate were resolved by SDS-PAGE and immunoblotting with the indicated antibodies. The expression levels of beta actin were used to ensure equal loading. Above: Representative Western blots, Below: Western blot quantification. Protein levels were normalized to beta actin, and were normalized to the level of untreated control (dashed line). One way-ANOVA with Bonferroni post-test was used to calculate statistics: * p

    Techniques Used: SDS Page, Expressing, Western Blot

    23) Product Images from "CyclinA1 Expression and Paclitaxel Resistance in Human Ovarian Cancer Cells"

    Article Title: CyclinA1 Expression and Paclitaxel Resistance in Human Ovarian Cancer Cells

    Journal: European journal of cancer (Oxford, England : 1990)

    doi: 10.1016/j.ejca.2016.08.007

    Apoptosis assay and senescence-associated beta-galactosidase assay TOV112D-CCNA1 ( A ) and TOV112D-pCDNA4 ( B ) were treated with various concentrations of paclitaxel for 48 hours either in the induced (1μg/ml tetracycline), or non-induced (without tetracycline) conditions, respectively, and then subject to apoptosis assay as described in the Materials and Methods. ( C ) Cytochemical detection of lysosomal beta-galactosidase activity at pH 4.0 (upper panel) and senescence-associated beta-galactosidase activity at pH 6.0 (bottom panel) of control (TOV112D-pcDNA4) cells, and cylin A1-induced (TOV112D-CCNA1) cells. The senescence-associated beta-galactosidase stained TOV112D-CCNA1 cells are marked by arrow blocks.
    Figure Legend Snippet: Apoptosis assay and senescence-associated beta-galactosidase assay TOV112D-CCNA1 ( A ) and TOV112D-pCDNA4 ( B ) were treated with various concentrations of paclitaxel for 48 hours either in the induced (1μg/ml tetracycline), or non-induced (without tetracycline) conditions, respectively, and then subject to apoptosis assay as described in the Materials and Methods. ( C ) Cytochemical detection of lysosomal beta-galactosidase activity at pH 4.0 (upper panel) and senescence-associated beta-galactosidase activity at pH 6.0 (bottom panel) of control (TOV112D-pcDNA4) cells, and cylin A1-induced (TOV112D-CCNA1) cells. The senescence-associated beta-galactosidase stained TOV112D-CCNA1 cells are marked by arrow blocks.

    Techniques Used: Apoptosis Assay, β-Gal Assay, Activity Assay, Staining

    24) Product Images from "t10c12 Conjugated Linoleic Acid Suppresses HER2 Protein and Enhances Apoptosis in SKBr3 Breast Cancer Cells: Possible Role of COX2"

    Article Title: t10c12 Conjugated Linoleic Acid Suppresses HER2 Protein and Enhances Apoptosis in SKBr3 Breast Cancer Cells: Possible Role of COX2

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0005342

    t10c12 CLA reduces nuclear p65 in SKBr3 cells. (A) Western blots of NFκB p65 in nuclear extracts. Cells were plated in 6-well plates, 3 wells per treatment. Cells from 3 wells were pulled and nuclear extract was obtained as described in Materials and Methods . 10 µg of nuclear extract was loaded into 10% gels. Gel electrophoresis and immunoblots were performed as described in Materials and Methods . Densitometry of bands was performed using Scion Image software Alpha 4.0.3.2. Protein expression of p65 was normalized to total protein and compared to levels in untreated cells (vehicle only) (* = 0.05). Values represent means +/− std error relative to vehicle control from 3 independent experiments. (B) Immunofluorescence of p65. Cells were treated as above. Immunoflourescence was performed as described in Materials and Methods . Top panel: vehicle; Middle pane: 40 µM CLA; Bottom panel: 80 µM CLA. Images were obtained using Delta Vision deconvolution microscope with SoftwoRX 3.5.0 software at 40× and optimized using Adobe PhotoShop CS2 version 9.0.2. (C) Western blot of total and phosphorylated IkappaB protein. Cells were plated and treated as above. 25 µg of whole cell lysate were loaded into 10% gels. Expression levels of IκB proteins were normalized to beta actin and compared to levels in untreated cells (vehicle only). Values represent means +/− std dev from 3 independent experiments (* = 0.05). Gel electrophoresis and immunoblots were performed as described in Materials and Methods . Densitometry of bands was performed using Scion Image software Alpha 4.0.3.2.
    Figure Legend Snippet: t10c12 CLA reduces nuclear p65 in SKBr3 cells. (A) Western blots of NFκB p65 in nuclear extracts. Cells were plated in 6-well plates, 3 wells per treatment. Cells from 3 wells were pulled and nuclear extract was obtained as described in Materials and Methods . 10 µg of nuclear extract was loaded into 10% gels. Gel electrophoresis and immunoblots were performed as described in Materials and Methods . Densitometry of bands was performed using Scion Image software Alpha 4.0.3.2. Protein expression of p65 was normalized to total protein and compared to levels in untreated cells (vehicle only) (* = 0.05). Values represent means +/− std error relative to vehicle control from 3 independent experiments. (B) Immunofluorescence of p65. Cells were treated as above. Immunoflourescence was performed as described in Materials and Methods . Top panel: vehicle; Middle pane: 40 µM CLA; Bottom panel: 80 µM CLA. Images were obtained using Delta Vision deconvolution microscope with SoftwoRX 3.5.0 software at 40× and optimized using Adobe PhotoShop CS2 version 9.0.2. (C) Western blot of total and phosphorylated IkappaB protein. Cells were plated and treated as above. 25 µg of whole cell lysate were loaded into 10% gels. Expression levels of IκB proteins were normalized to beta actin and compared to levels in untreated cells (vehicle only). Values represent means +/− std dev from 3 independent experiments (* = 0.05). Gel electrophoresis and immunoblots were performed as described in Materials and Methods . Densitometry of bands was performed using Scion Image software Alpha 4.0.3.2.

    Techniques Used: Western Blot, Nucleic Acid Electrophoresis, Software, Expressing, Immunofluorescence, Microscopy

    t10c12 CLA reduces HER2 protein in SK-Br3 cells. (A) Representative western blot of total HER2 protein in response to 24 hr treatment with t10c12 CLA. Cells were plated in 6-well plates, 3 wells per treatment. Cells from 3 wells were pulled and total protein was extracted as described in Materials and Methods . 25 µg of whole cell lysate were loaded into 8% gels. Densitometry of bands was performed using Scion Image software Alpha 4.0.3.2. Expression of HER2 was normalized to beta actin and compared to vehicle treatment. Values represent the mean +/− std error relative to vehicle from 5 independent experiments. (B) Immunofluoresnce of HER2 protein in SK-Br3 cells following 24 hr treatment with 40 µM (middle panel), 80 µM (right panel) or vehicle (left panel). Cells were treated and immunofluorescence was performed as described in Materials and Methods . Images were obtained using Delta Vision deconvolution microscope with SoftwoRX 3.5.0 software at 40× and optimized using Adobe PhotoShop CS2 version 9.0.2.
    Figure Legend Snippet: t10c12 CLA reduces HER2 protein in SK-Br3 cells. (A) Representative western blot of total HER2 protein in response to 24 hr treatment with t10c12 CLA. Cells were plated in 6-well plates, 3 wells per treatment. Cells from 3 wells were pulled and total protein was extracted as described in Materials and Methods . 25 µg of whole cell lysate were loaded into 8% gels. Densitometry of bands was performed using Scion Image software Alpha 4.0.3.2. Expression of HER2 was normalized to beta actin and compared to vehicle treatment. Values represent the mean +/− std error relative to vehicle from 5 independent experiments. (B) Immunofluoresnce of HER2 protein in SK-Br3 cells following 24 hr treatment with 40 µM (middle panel), 80 µM (right panel) or vehicle (left panel). Cells were treated and immunofluorescence was performed as described in Materials and Methods . Images were obtained using Delta Vision deconvolution microscope with SoftwoRX 3.5.0 software at 40× and optimized using Adobe PhotoShop CS2 version 9.0.2.

    Techniques Used: Western Blot, Software, Expressing, Immunofluorescence, Microscopy

    25) Product Images from "Human immunodeficiency virus Tat impairs mitochondrial fission in neurons"

    Article Title: Human immunodeficiency virus Tat impairs mitochondrial fission in neurons

    Journal: Cell Death Discovery

    doi: 10.1038/s41420-017-0013-6

    Tat promotes a time-dependent increase in Drp1, but not Mfn2 levels. Drp1 and Mfns protein levels were determined by western blot analysis in control and Tat-treated neurons. a Representative western blot analysis of cortical neuronal lysates probed with a Drp1 antibody. Blots were reprobed with beta-actin antibody. b Semi-quantification of Drp1 levels was done by densitometric analysis of the 82 kDa immunoreactive band normalized by the beta-actin (42 kDa) immunoreactivity. c Representative western blot analysis of cortical neuronal lysates probed with a Mfn2 antibody. d Semi-quantification of Mfn2 levels was done by densitometric analysis of the 86 kDa immunoreactive band normalized by beta-actin immunoreacivity. Data are the mean ± SEM of three independent experiments, normalized to control. * p
    Figure Legend Snippet: Tat promotes a time-dependent increase in Drp1, but not Mfn2 levels. Drp1 and Mfns protein levels were determined by western blot analysis in control and Tat-treated neurons. a Representative western blot analysis of cortical neuronal lysates probed with a Drp1 antibody. Blots were reprobed with beta-actin antibody. b Semi-quantification of Drp1 levels was done by densitometric analysis of the 82 kDa immunoreactive band normalized by the beta-actin (42 kDa) immunoreactivity. c Representative western blot analysis of cortical neuronal lysates probed with a Mfn2 antibody. d Semi-quantification of Mfn2 levels was done by densitometric analysis of the 86 kDa immunoreactive band normalized by beta-actin immunoreacivity. Data are the mean ± SEM of three independent experiments, normalized to control. * p

    Techniques Used: Western Blot

    26) Product Images from "Loss of Igfbp7 Causes Precocious Involution in Lactating Mouse Mammary Gland"

    Article Title: Loss of Igfbp7 Causes Precocious Involution in Lactating Mouse Mammary Gland

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0087858

    The Igfbp7 −/− transcriptome is enriched for involution-associate gene signature. (A) The transcriptome profiles of the Wild-Type (WT) and the Igfbp7 −/− glands on the lactation day 3 were analyzed using the RNA-Seq technique. Differentially expressed transcripts were identified by imposing a minimum of 1.5 fold change in the transcript expression level and a P-value of 0.05 or lower and are depicted on a Volcano plot. (B and C) The differentially expressed transcripts in the Igfbp7 −/− mammary epithelial cells (MECs) are categorized into the different KEGG Signaling Pathways using the DAVID Bioinformatics Resources. The fold enrichment scores are calculated based on the number of differentially regulated genes that belong to a particular KEGG signaling pathway out of the total gene set. The Benjamini statistics is used to identify the statistically significant gene enrichment for specific KEGG signaling pathways. Using this analysis the down regulated transcripts were found to be enriched in extracellular matrix receptor interaction, focal adhesion and cell adhesion molecules. The up regulated transcripts in the Igfpb7 −/− MECs showed enrichment for the WNT, TGF beta and adherens junction proteins. (D) To examine if the differentially regulated transcripts in the Igfbp7 −/− MECs show enrichment for involution-related genes, we first obtained a common involution gene signature by comparing 3 publically available transcriptome profile of involuting glands. Set 1 is from [35] , Set 2 is from [34] , Set 3 is from [33] . The Ven diagram depicts a 72 gene-set that is common among all 3 data sets.
    Figure Legend Snippet: The Igfbp7 −/− transcriptome is enriched for involution-associate gene signature. (A) The transcriptome profiles of the Wild-Type (WT) and the Igfbp7 −/− glands on the lactation day 3 were analyzed using the RNA-Seq technique. Differentially expressed transcripts were identified by imposing a minimum of 1.5 fold change in the transcript expression level and a P-value of 0.05 or lower and are depicted on a Volcano plot. (B and C) The differentially expressed transcripts in the Igfbp7 −/− mammary epithelial cells (MECs) are categorized into the different KEGG Signaling Pathways using the DAVID Bioinformatics Resources. The fold enrichment scores are calculated based on the number of differentially regulated genes that belong to a particular KEGG signaling pathway out of the total gene set. The Benjamini statistics is used to identify the statistically significant gene enrichment for specific KEGG signaling pathways. Using this analysis the down regulated transcripts were found to be enriched in extracellular matrix receptor interaction, focal adhesion and cell adhesion molecules. The up regulated transcripts in the Igfpb7 −/− MECs showed enrichment for the WNT, TGF beta and adherens junction proteins. (D) To examine if the differentially regulated transcripts in the Igfbp7 −/− MECs show enrichment for involution-related genes, we first obtained a common involution gene signature by comparing 3 publically available transcriptome profile of involuting glands. Set 1 is from [35] , Set 2 is from [34] , Set 3 is from [33] . The Ven diagram depicts a 72 gene-set that is common among all 3 data sets.

    Techniques Used: RNA Sequencing Assay, Expressing

    Lactating Igfbp7 −/− glands exhibit accelerated involution. To determine if Igfbp7 −/− glands exhibit molecular changes that are the hallmark of involution process we prepared protein extracts from the Wild Type (WT) or the Igfbp7 −/− glands on lactation day 3 (WT LD3, KO LD3 respectively) or from WT lactating glands were weaned for 5 days (WT Inv D5) to induce post-lactational involution. The expression of Stat5a, Stat3, phospho Stat3 (pStat3), AKT, pAKT, Igfbp5, and IGF-1R proteins was determined by Western Blots. (A, C–D) Representative Western Blots are shown. The protein expression levels for Stat5a and Stat3, AKT, Igfbp5, and IGF-1R have been normalized to beta actin expression while the expression of pStat3, pAKT have been normalized to total corresponding protein expression in each sample. The bar graphs show the average expression obtained from of 3 independent protein extracts and the statistical significance was calculated based on two-tailed t-test (*P =
    Figure Legend Snippet: Lactating Igfbp7 −/− glands exhibit accelerated involution. To determine if Igfbp7 −/− glands exhibit molecular changes that are the hallmark of involution process we prepared protein extracts from the Wild Type (WT) or the Igfbp7 −/− glands on lactation day 3 (WT LD3, KO LD3 respectively) or from WT lactating glands were weaned for 5 days (WT Inv D5) to induce post-lactational involution. The expression of Stat5a, Stat3, phospho Stat3 (pStat3), AKT, pAKT, Igfbp5, and IGF-1R proteins was determined by Western Blots. (A, C–D) Representative Western Blots are shown. The protein expression levels for Stat5a and Stat3, AKT, Igfbp5, and IGF-1R have been normalized to beta actin expression while the expression of pStat3, pAKT have been normalized to total corresponding protein expression in each sample. The bar graphs show the average expression obtained from of 3 independent protein extracts and the statistical significance was calculated based on two-tailed t-test (*P =

    Techniques Used: Expressing, Western Blot, Two Tailed Test

    27) Product Images from "Characterization of Zebrafish Models of Marinesco-Sjögren Syndrome"

    Article Title: Characterization of Zebrafish Models of Marinesco-Sjögren Syndrome

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0165563

    Immunohistochemistry of skeletal muscle tissue of morpholino-injected fish. Immunostaining of CMO injected fish (A, B) and sil1 morphant (MO2) injected fish (C, D) with antibodies against beta-dystroglycan (beta-DG) (A, C) and myosin heavy chain (MHC) (B, D). Beta-dystroglycan expression at the myosepta of MO2 injected 4 dpf embryos is misshapen and has a less clear v-shaped structure. Staining with anti-MHC indicated that formation of myofibers is disturbed in MO2. Arrowheads indicate the disturbance of myosepta in MO2-injected fish. Bar: 100 μm. Arrows indicate the abnormal structure of myofibers.
    Figure Legend Snippet: Immunohistochemistry of skeletal muscle tissue of morpholino-injected fish. Immunostaining of CMO injected fish (A, B) and sil1 morphant (MO2) injected fish (C, D) with antibodies against beta-dystroglycan (beta-DG) (A, C) and myosin heavy chain (MHC) (B, D). Beta-dystroglycan expression at the myosepta of MO2 injected 4 dpf embryos is misshapen and has a less clear v-shaped structure. Staining with anti-MHC indicated that formation of myofibers is disturbed in MO2. Arrowheads indicate the disturbance of myosepta in MO2-injected fish. Bar: 100 μm. Arrows indicate the abnormal structure of myofibers.

    Techniques Used: Immunohistochemistry, Injection, Fluorescence In Situ Hybridization, Immunostaining, Expressing, Staining

    Protein expression analysis of sil1-morpholino injected fish. Western blot analysis of control morpholino (CMO) and sil1-morpholino2 injected fish (MO). The protein amounts of BiP, lipidated form of LC3 (LC3-II), and activated caspase 3 are significantly increased in sil1 morphant embryos compared to those of CMO-injected embryos. A: Immunoblot with anti-BiP, LC3-II, activated-Caspase 3, and beta-actin. B: Relative expression level is analyzed via immunoblot (*p = 0.0226, ** p = 0.000102 and *** p = 0.000705, versus CMO, Student’s t-test, n = 3).
    Figure Legend Snippet: Protein expression analysis of sil1-morpholino injected fish. Western blot analysis of control morpholino (CMO) and sil1-morpholino2 injected fish (MO). The protein amounts of BiP, lipidated form of LC3 (LC3-II), and activated caspase 3 are significantly increased in sil1 morphant embryos compared to those of CMO-injected embryos. A: Immunoblot with anti-BiP, LC3-II, activated-Caspase 3, and beta-actin. B: Relative expression level is analyzed via immunoblot (*p = 0.0226, ** p = 0.000102 and *** p = 0.000705, versus CMO, Student’s t-test, n = 3).

    Techniques Used: Expressing, Injection, Fluorescence In Situ Hybridization, Western Blot

    28) Product Images from "Alpha-synuclein deficiency leads to increased glyoxalase I expression and glycation stress"

    Article Title: Alpha-synuclein deficiency leads to increased glyoxalase I expression and glycation stress

    Journal: Cellular and Molecular Life Sciences

    doi: 10.1007/s00018-010-0483-7

    Elevation of GLO1 protein levels in striatum of alpha-synuclein-deficient mice. a Immunoblot analysis revealed b significant elevated GLO1 protein levels (3.26 ± 0.32-fold; n = 5/genotype, p = 0.0003) in the striatum of KO mice at old age (19 months) in comparison with corresponding WT controls. Protein expression levels of beta-actin were used for normalization of GLO1 protein levels and to confirm equal protein loading. Data are presented as mean ± SD and significant differences were highlighted with asterisks (*** p
    Figure Legend Snippet: Elevation of GLO1 protein levels in striatum of alpha-synuclein-deficient mice. a Immunoblot analysis revealed b significant elevated GLO1 protein levels (3.26 ± 0.32-fold; n = 5/genotype, p = 0.0003) in the striatum of KO mice at old age (19 months) in comparison with corresponding WT controls. Protein expression levels of beta-actin were used for normalization of GLO1 protein levels and to confirm equal protein loading. Data are presented as mean ± SD and significant differences were highlighted with asterisks (*** p

    Techniques Used: Mouse Assay, Expressing

    29) Product Images from "Reduced migration of MLH1 deficient colon cancer cells depends on SPTAN1"

    Article Title: Reduced migration of MLH1 deficient colon cancer cells depends on SPTAN1

    Journal: Molecular Cancer

    doi: 10.1186/1476-4598-13-11

    Influence of MLH1 knock down by siRNA treatment. Direct connection of MLH1 on SPTAN1 was studied in detail by comparison of untreated with transiently MLH1-specific siRNA1 (A) and siRNA2 (B) and control siRNA transfected different MLH1 proficient cell lines. Western blot analysis of MLH1 and SPTAN1 protein levels was performed after 48 h, controlled by beta-Actin detection after. In parallel, mRNA levels of MLH1 and SPTAN1 were determined by TaqMan-analysis with pairs of MLH1-, SPTAN1- or GAPDH-specific primers and MLH1-, SPTAN1- or GAPDH-specific probes, respectively, after treatment with MLH1-specific siRNA1 (C) and siRNA2 (D) . The data show that siRNA knock down of MLH1 led to impaired SPTAN1 expression and partly to reduced mRNA level of SPTAN1. Graphs indicate the results (mean ± S.D.) of at least three independent experiments.
    Figure Legend Snippet: Influence of MLH1 knock down by siRNA treatment. Direct connection of MLH1 on SPTAN1 was studied in detail by comparison of untreated with transiently MLH1-specific siRNA1 (A) and siRNA2 (B) and control siRNA transfected different MLH1 proficient cell lines. Western blot analysis of MLH1 and SPTAN1 protein levels was performed after 48 h, controlled by beta-Actin detection after. In parallel, mRNA levels of MLH1 and SPTAN1 were determined by TaqMan-analysis with pairs of MLH1-, SPTAN1- or GAPDH-specific primers and MLH1-, SPTAN1- or GAPDH-specific probes, respectively, after treatment with MLH1-specific siRNA1 (C) and siRNA2 (D) . The data show that siRNA knock down of MLH1 led to impaired SPTAN1 expression and partly to reduced mRNA level of SPTAN1. Graphs indicate the results (mean ± S.D.) of at least three independent experiments.

    Techniques Used: Transfection, Western Blot, Expressing

    MLH1 deficient cell lines show reduced SPTAN1 expression. To determine a potential influence of MLH1 on SPTAN1 expression different MLH1 proficient and MLH1 deficient cell lines were analyzed by (A) Western blotting using anti-MLH1 or anti-SPTAN1, respectively, controlled by beta-Actin detection. (B) Amounts of SPTAN1 were assessed by measuring the signal intensities of protein bands with Multi Gauge V3.2 software. Graphs indicate the results (mean ± S.D.) of at least four independent experiments. Comparability was facilitated by setting SPTAN1 levels of HeLa (lane 1) as 100%. Shared identity of MLH1 proficient with MLH1 deficient sister cell lines is assigned by grey arrows.
    Figure Legend Snippet: MLH1 deficient cell lines show reduced SPTAN1 expression. To determine a potential influence of MLH1 on SPTAN1 expression different MLH1 proficient and MLH1 deficient cell lines were analyzed by (A) Western blotting using anti-MLH1 or anti-SPTAN1, respectively, controlled by beta-Actin detection. (B) Amounts of SPTAN1 were assessed by measuring the signal intensities of protein bands with Multi Gauge V3.2 software. Graphs indicate the results (mean ± S.D.) of at least four independent experiments. Comparability was facilitated by setting SPTAN1 levels of HeLa (lane 1) as 100%. Shared identity of MLH1 proficient with MLH1 deficient sister cell lines is assigned by grey arrows.

    Techniques Used: Expressing, Western Blot, Software

    Nuclear and cytoplasmic SPTAN1 analysis. SPTAN1 was described to be involved in DNA repair processes. To determine cellular localization of SPTAN1, nuclear (N) and cytoplasmic (C) protein fractions of SPTAN1 overexpressing or corresponding untransfected HEK293 and HEK293T cells, cells were harvested and analyzed by Western blotting. The efficiency of the fractionation was verified by staining for Lamin B as a nuclear marker and gamma Adaptin as a cytoplasmic marker. In parallel, beta-Actin detection served for quantitative control.
    Figure Legend Snippet: Nuclear and cytoplasmic SPTAN1 analysis. SPTAN1 was described to be involved in DNA repair processes. To determine cellular localization of SPTAN1, nuclear (N) and cytoplasmic (C) protein fractions of SPTAN1 overexpressing or corresponding untransfected HEK293 and HEK293T cells, cells were harvested and analyzed by Western blotting. The efficiency of the fractionation was verified by staining for Lamin B as a nuclear marker and gamma Adaptin as a cytoplasmic marker. In parallel, beta-Actin detection served for quantitative control.

    Techniques Used: Western Blot, Fractionation, Staining, Marker

    30) Product Images from "Myeloid antigens in childhood lymphoblastic leukemia:clinical data point to regulation of CD66c distinct from other myeloid antigens"

    Article Title: Myeloid antigens in childhood lymphoblastic leukemia:clinical data point to regulation of CD66c distinct from other myeloid antigens

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-5-38

    Transcription of CEACAM6 versus surface CD66c expression on sorted cells. FACSsorted CD66c surface negative (CD66c neg ) or positive (CD66c pos ) ALL lymphoblasts, five patients with heterogeneous CD66c expression were sorted into both CD66c negative and CD66c positive fraction (lines connect sorted fractions from the same specimen). Mann-Whitney test was used to compare groups (n = 32). CEACAM6n value is normalized to beta-2-microglobulin (see Methods).
    Figure Legend Snippet: Transcription of CEACAM6 versus surface CD66c expression on sorted cells. FACSsorted CD66c surface negative (CD66c neg ) or positive (CD66c pos ) ALL lymphoblasts, five patients with heterogeneous CD66c expression were sorted into both CD66c negative and CD66c positive fraction (lines connect sorted fractions from the same specimen). Mann-Whitney test was used to compare groups (n = 32). CEACAM6n value is normalized to beta-2-microglobulin (see Methods).

    Techniques Used: Expressing, MANN-WHITNEY

    31) Product Images from "Human immunodeficiency virus Tat impairs mitochondrial fission in neurons"

    Article Title: Human immunodeficiency virus Tat impairs mitochondrial fission in neurons

    Journal: Cell Death Discovery

    doi: 10.1038/s41420-017-0013-6

    Tat promotes a time-dependent increase in Drp1, but not Mfn2 levels. Drp1 and Mfns protein levels were determined by western blot analysis in control and Tat-treated neurons. a Representative western blot analysis of cortical neuronal lysates probed with a Drp1 antibody. Blots were reprobed with beta-actin antibody. b Semi-quantification of Drp1 levels was done by densitometric analysis of the 82 kDa immunoreactive band normalized by the beta-actin (42 kDa) immunoreactivity. c Representative western blot analysis of cortical neuronal lysates probed with a Mfn2 antibody. d Semi-quantification of Mfn2 levels was done by densitometric analysis of the 86 kDa immunoreactive band normalized by beta-actin immunoreacivity. Data are the mean ± SEM of three independent experiments, normalized to control. * p
    Figure Legend Snippet: Tat promotes a time-dependent increase in Drp1, but not Mfn2 levels. Drp1 and Mfns protein levels were determined by western blot analysis in control and Tat-treated neurons. a Representative western blot analysis of cortical neuronal lysates probed with a Drp1 antibody. Blots were reprobed with beta-actin antibody. b Semi-quantification of Drp1 levels was done by densitometric analysis of the 82 kDa immunoreactive band normalized by the beta-actin (42 kDa) immunoreactivity. c Representative western blot analysis of cortical neuronal lysates probed with a Mfn2 antibody. d Semi-quantification of Mfn2 levels was done by densitometric analysis of the 86 kDa immunoreactive band normalized by beta-actin immunoreacivity. Data are the mean ± SEM of three independent experiments, normalized to control. * p

    Techniques Used: Western Blot

    32) Product Images from "Inflammatory and cytotoxic effects of bifenthrin in primary microglia and organotypic hippocampal slice cultures"

    Article Title: Inflammatory and cytotoxic effects of bifenthrin in primary microglia and organotypic hippocampal slice cultures

    Journal: Journal of Neuroinflammation

    doi: 10.1186/s12974-018-1198-1

    BF exposure increases the expression of pro-inflammatory markers in primary microglia cells. Microglial cells were exposed to different concentrations of BF (1–20 μM) for 24 h. Gene expression of TNF-alpha ( a ), IL-6 ( b ), COX-2 ( c ), and mPGES-1 ( d ) was analyzed by real-time quantitative PCR. GAPDH was used as an internal control for normalization, and data were quantified by using the comparative cycle threshold Ct method. Similarly, cells were treated with BF and thereafter incubated with or without LPS (100 ng/mL) as a positive control (black column) for 24 h. Whole cell lysates were subjected to Western blot for COX-2 ( e , f ), mPGES-1 ( g , h ), and beta-actin. To confirm equal sample loading, beta-actin was used for normalization. Data are presented as percentage control of DMSO. Statistical analyses were carried out by using one-way ANOVA followed by post hoc Student–Newman–Keuls test. Results are expressed as means ± SEM of three independent experiments. *p
    Figure Legend Snippet: BF exposure increases the expression of pro-inflammatory markers in primary microglia cells. Microglial cells were exposed to different concentrations of BF (1–20 μM) for 24 h. Gene expression of TNF-alpha ( a ), IL-6 ( b ), COX-2 ( c ), and mPGES-1 ( d ) was analyzed by real-time quantitative PCR. GAPDH was used as an internal control for normalization, and data were quantified by using the comparative cycle threshold Ct method. Similarly, cells were treated with BF and thereafter incubated with or without LPS (100 ng/mL) as a positive control (black column) for 24 h. Whole cell lysates were subjected to Western blot for COX-2 ( e , f ), mPGES-1 ( g , h ), and beta-actin. To confirm equal sample loading, beta-actin was used for normalization. Data are presented as percentage control of DMSO. Statistical analyses were carried out by using one-way ANOVA followed by post hoc Student–Newman–Keuls test. Results are expressed as means ± SEM of three independent experiments. *p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Incubation, Positive Control, Western Blot

    BF exposure increases the expression of Nrf-2 and NF-kappaB in primary microglia. Microglial cells were exposed to different concentrations of BF (1–20 μM) for 24 h. Gene expression of Nrf-2 ( a ) and NF-kappaBp65 ( b ) was analyzed by real-time quantitative PCR. GAPDH was used as an internal control for normalization, and data were quantified by using the comparative cycle threshold Ct method. Similarly, cells were treated with BF and thereafter incubated with or without LPS (100 ng/mL) as a positive control (black column) for 24 h. Whole cell lysates were subjected to Western blot for Nrf-2 ( c , d ), NF-kappaBp65 ( e , f ), and beta-actin. Data are presented as percentage control of DMSO. Statistical analyses were carried out by using one-way ANOVA followed by post hoc Student–Newman–Keuls test. Results are expressed as means ± SEM of three independent experiments. * p
    Figure Legend Snippet: BF exposure increases the expression of Nrf-2 and NF-kappaB in primary microglia. Microglial cells were exposed to different concentrations of BF (1–20 μM) for 24 h. Gene expression of Nrf-2 ( a ) and NF-kappaBp65 ( b ) was analyzed by real-time quantitative PCR. GAPDH was used as an internal control for normalization, and data were quantified by using the comparative cycle threshold Ct method. Similarly, cells were treated with BF and thereafter incubated with or without LPS (100 ng/mL) as a positive control (black column) for 24 h. Whole cell lysates were subjected to Western blot for Nrf-2 ( c , d ), NF-kappaBp65 ( e , f ), and beta-actin. Data are presented as percentage control of DMSO. Statistical analyses were carried out by using one-way ANOVA followed by post hoc Student–Newman–Keuls test. Results are expressed as means ± SEM of three independent experiments. * p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Incubation, Positive Control, Western Blot

    33) Product Images from "Reciprocal and Autonomous Glucocorticoid and Androgen Receptor Activation in Salivary Duct Carcinoma"

    Article Title: Reciprocal and Autonomous Glucocorticoid and Androgen Receptor Activation in Salivary Duct Carcinoma

    Journal: Clinical cancer research : an official journal of the American Association for Cancer Research

    doi: 10.1158/1078-0432.CCR-19-1603

    GR expression and its association with poor survival in SDCs. (A) Glucocorticoid receptor (GR) and androgen receptor (AR) protein expression by immunohistochemical (IHC) analysis. Case SDC-09 demonstrated high GR (nuclear staining intensity in tumor cells is higher than in stroma cells) and were AR negative. Case SDC-17 cells demonstrated normal GR expression levels, which were the same as those in stroma cells, and were AR positive. Case SDC-66 showed both loss of GR (stroma is positive GR) and AR in tumor cells. (B) GR and AR expression profile in SDCs. (C) Western blot analysis of GR and AR protein expression in selected SDCs. High GR proteins by IHC analysis corresponded to stronger GR expression in SDC tissue than in normal salivary gland tissues. Beta-actin (ACTB) was used as the loading control. Status of AR and GR by IHC analysis, shown at bottom; positive (P) and negative (N). Status of GR by IHC analysis; high (H), constitutive (C), and lost (L). (D) Kaplan-Meier analysis revealed a significant correlation between high GR expression and a low overall survival rate (upper panel, P = 0.046, log-rank test). Of note, there was a trend towards a higher overall survival rate in patients with constitutive GR expression and loss of GR expression (lower panel).
    Figure Legend Snippet: GR expression and its association with poor survival in SDCs. (A) Glucocorticoid receptor (GR) and androgen receptor (AR) protein expression by immunohistochemical (IHC) analysis. Case SDC-09 demonstrated high GR (nuclear staining intensity in tumor cells is higher than in stroma cells) and were AR negative. Case SDC-17 cells demonstrated normal GR expression levels, which were the same as those in stroma cells, and were AR positive. Case SDC-66 showed both loss of GR (stroma is positive GR) and AR in tumor cells. (B) GR and AR expression profile in SDCs. (C) Western blot analysis of GR and AR protein expression in selected SDCs. High GR proteins by IHC analysis corresponded to stronger GR expression in SDC tissue than in normal salivary gland tissues. Beta-actin (ACTB) was used as the loading control. Status of AR and GR by IHC analysis, shown at bottom; positive (P) and negative (N). Status of GR by IHC analysis; high (H), constitutive (C), and lost (L). (D) Kaplan-Meier analysis revealed a significant correlation between high GR expression and a low overall survival rate (upper panel, P = 0.046, log-rank test). Of note, there was a trend towards a higher overall survival rate in patients with constitutive GR expression and loss of GR expression (lower panel).

    Techniques Used: Expressing, Immunohistochemistry, Staining, Western Blot

    GR inhibition did not affect AR expression. (A) Protein GR and AR expression in SDC cells. All three cell lines expressed GR. Of note, RET981 cells expressed relatively high GR. Beta-actin (ACTB) was used as a loading control. (B) GR siRNAs inhibited cell proliferation in all SDC cells after 72 hours. Error bars in all graphs represent standard deviation. Asterisks (*) represent significant differences in proliferation activity compared with the control ( P
    Figure Legend Snippet: GR inhibition did not affect AR expression. (A) Protein GR and AR expression in SDC cells. All three cell lines expressed GR. Of note, RET981 cells expressed relatively high GR. Beta-actin (ACTB) was used as a loading control. (B) GR siRNAs inhibited cell proliferation in all SDC cells after 72 hours. Error bars in all graphs represent standard deviation. Asterisks (*) represent significant differences in proliferation activity compared with the control ( P

    Techniques Used: Inhibition, Expressing, Standard Deviation, Activity Assay

    34) Product Images from "MICAL2 is a novel human cancer gene controlling mesenchymal to epithelial transition involved in cancer growth and invasion"

    Article Title: MICAL2 is a novel human cancer gene controlling mesenchymal to epithelial transition involved in cancer growth and invasion

    Journal: Oncotarget

    doi:

    Abating MICAL2 in 786-O kidney cancer cells in vitro induces MET ( A ), Morphological analysis in light-transmitted microscopy. Scale bar: 100 μm. ( B ), F-actin staining. IF of: E-cadherin, ZO-1, catenin-beta. In KD2 and KD14 cells, these three markers relocated at cell perimeters (as in epithelial cells), rather than being diffusedly cytoplasmic. IF of vimentin with Vim13.2 antibody. In MIC2-KD cells it decorated short and disorganized filaments instead of long meshes observed in control cells. Scale bar of all IF images: 20 μm. Inset magnification: 3x. ( C ), Vimentin epitope recognized by V9 antibody was undetectable in KD2 and KD14 cells. ( D ), Agarose gel run of SNAI1, SNAI2, 18s5 RT-PCR products. ( E ), KD14 cells transfected with HA-MICAL2 cDNA. The elongated phenotype showed dose-dependency on HA-MICAL2 expression. One-way Anova test and Tukey's Multiple Comparison post-hoc test. In all graph bars, horizontal lines denote mean and SEM. N.s: non significant. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001). ( F ), MIC2-KD14 cells expressing HA-MICAL2 (shown by anti-HA immunostaining) recovered a mesenchymal-like phenotype with concomitant F-actin redistribution.
    Figure Legend Snippet: Abating MICAL2 in 786-O kidney cancer cells in vitro induces MET ( A ), Morphological analysis in light-transmitted microscopy. Scale bar: 100 μm. ( B ), F-actin staining. IF of: E-cadherin, ZO-1, catenin-beta. In KD2 and KD14 cells, these three markers relocated at cell perimeters (as in epithelial cells), rather than being diffusedly cytoplasmic. IF of vimentin with Vim13.2 antibody. In MIC2-KD cells it decorated short and disorganized filaments instead of long meshes observed in control cells. Scale bar of all IF images: 20 μm. Inset magnification: 3x. ( C ), Vimentin epitope recognized by V9 antibody was undetectable in KD2 and KD14 cells. ( D ), Agarose gel run of SNAI1, SNAI2, 18s5 RT-PCR products. ( E ), KD14 cells transfected with HA-MICAL2 cDNA. The elongated phenotype showed dose-dependency on HA-MICAL2 expression. One-way Anova test and Tukey's Multiple Comparison post-hoc test. In all graph bars, horizontal lines denote mean and SEM. N.s: non significant. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001). ( F ), MIC2-KD14 cells expressing HA-MICAL2 (shown by anti-HA immunostaining) recovered a mesenchymal-like phenotype with concomitant F-actin redistribution.

    Techniques Used: In Vitro, Microscopy, Staining, Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Transfection, Expressing, Immunostaining

    35) Product Images from "Semaphorin4A Is Cytotoxic to Oligodendrocytes and Is Elevated in Microglia and Multiple Sclerosis"

    Article Title: Semaphorin4A Is Cytotoxic to Oligodendrocytes and Is Elevated in Microglia and Multiple Sclerosis

    Journal: ASN NEURO

    doi: 10.1177/1759091415587502

    Sema4A is increased after microglial activation with LPS. (a and b) Sema4A is expressed by rat primary microglia and is increased upon activation with 0.8 µg/mL LPS for 24 hr. Hft is also increased after LPS treatment, indicating activation of the microglia. Data are representative of three independent experiments, completed in triplicate. Protein quantification is relative to beta-actin loading control. Values represent average ± SEM . Student’s t test was used to determine significance. LPS = lipopolysaccharide; Hft = H-ferritin. * p
    Figure Legend Snippet: Sema4A is increased after microglial activation with LPS. (a and b) Sema4A is expressed by rat primary microglia and is increased upon activation with 0.8 µg/mL LPS for 24 hr. Hft is also increased after LPS treatment, indicating activation of the microglia. Data are representative of three independent experiments, completed in triplicate. Protein quantification is relative to beta-actin loading control. Values represent average ± SEM . Student’s t test was used to determine significance. LPS = lipopolysaccharide; Hft = H-ferritin. * p

    Techniques Used: Activation Assay

    Microglial expression of Sema4A protein was increased after FAS induced iron loading while activated by LPS. Primary rat microglia cells were treated with increasing FAS or with 0.8 µg/mL LPS and increasing concentrations of FAS for 24 hr. (a) Sema4A is significantly increased after LPS alone. With the addition of LPS/FAS, there is a significant increase in Sema4A expression. (b) Hft expression is significantly increased after LPS alone. With the addition of LPS/FAS, expression is significantly increased. Protein quantification is relative to beta-actin loading control. Two-way analysis of variance with Bonferroni post hoc test was used to determine significance. Values represent average ± SEM . FAS = ferrous ammonium sulfate; Hft = H-ferritin. * p
    Figure Legend Snippet: Microglial expression of Sema4A protein was increased after FAS induced iron loading while activated by LPS. Primary rat microglia cells were treated with increasing FAS or with 0.8 µg/mL LPS and increasing concentrations of FAS for 24 hr. (a) Sema4A is significantly increased after LPS alone. With the addition of LPS/FAS, there is a significant increase in Sema4A expression. (b) Hft expression is significantly increased after LPS alone. With the addition of LPS/FAS, expression is significantly increased. Protein quantification is relative to beta-actin loading control. Two-way analysis of variance with Bonferroni post hoc test was used to determine significance. Values represent average ± SEM . FAS = ferrous ammonium sulfate; Hft = H-ferritin. * p

    Techniques Used: Expressing

    Microglial expression of Sema4A protein was decreased after DFO induced iron deficiency while activated by LPS. Primary rat microglia cells were treated with increasing DFO or with 0.8 µg/mL LPS and increasing concentrations of DFO for 24 hr. (a) Sema4A is significantly increased after LPS alone. With the addition of LPS/DFO, there is a significant decrease in Sema4A expression. (b) Hft expression is significantly increased after LPS alone. With the addition of LPS/DFO, Hft expression is significantly decreased. Protein quantification is relative to beta-actin loading control. Two-way analysis of variance with Bonferroni post hoc test was used to determine significance. Values represent average ± SEM . DFO = deferoxamine; LPS = lipopolysaccharide; Hft = H-ferritin. * p
    Figure Legend Snippet: Microglial expression of Sema4A protein was decreased after DFO induced iron deficiency while activated by LPS. Primary rat microglia cells were treated with increasing DFO or with 0.8 µg/mL LPS and increasing concentrations of DFO for 24 hr. (a) Sema4A is significantly increased after LPS alone. With the addition of LPS/DFO, there is a significant decrease in Sema4A expression. (b) Hft expression is significantly increased after LPS alone. With the addition of LPS/DFO, Hft expression is significantly decreased. Protein quantification is relative to beta-actin loading control. Two-way analysis of variance with Bonferroni post hoc test was used to determine significance. Values represent average ± SEM . DFO = deferoxamine; LPS = lipopolysaccharide; Hft = H-ferritin. * p

    Techniques Used: Expressing

    36) Product Images from "Pancreas-Specific Sirt1-Deficiency in Mice Compromises Beta-Cell Function without Development of Hyperglycemia"

    Article Title: Pancreas-Specific Sirt1-Deficiency in Mice Compromises Beta-Cell Function without Development of Hyperglycemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0128012

    Pdx1 Cre ; Sirt1 ex4F/F islets have reduced expression of Slc2a2/Glut2. A. Mitochondrial and fatty acid metabolism gene transcript levels analyzed by RT-qPCR in Pdx1 Cre and Pdx1 Cre ; Sirt1 ex4F/F isolated islets. Values are relative to the housekeeping gene. ( n ≥6). B. Endocrine transcription factors transcript levels analyzed by RT-qPCR in Pdx1 Cre and Pdx1 Cre ; Sirt1 ex4F/F isolated islets. Values are relative to the housekeeping gene. ( n ≥6). C. Beta-cell function gene transcript levels analyzed by RT-qPCR in Pdx1 Cre and Pdx1 Cre ; Sirt1 ex4F/F isolated islets. Values are relative to the housekeeping gene. ( n ≥6, * p
    Figure Legend Snippet: Pdx1 Cre ; Sirt1 ex4F/F islets have reduced expression of Slc2a2/Glut2. A. Mitochondrial and fatty acid metabolism gene transcript levels analyzed by RT-qPCR in Pdx1 Cre and Pdx1 Cre ; Sirt1 ex4F/F isolated islets. Values are relative to the housekeeping gene. ( n ≥6). B. Endocrine transcription factors transcript levels analyzed by RT-qPCR in Pdx1 Cre and Pdx1 Cre ; Sirt1 ex4F/F isolated islets. Values are relative to the housekeeping gene. ( n ≥6). C. Beta-cell function gene transcript levels analyzed by RT-qPCR in Pdx1 Cre and Pdx1 Cre ; Sirt1 ex4F/F isolated islets. Values are relative to the housekeeping gene. ( n ≥6, * p

    Techniques Used: Expressing, Quantitative RT-PCR, Isolation, Cell Function Assay

    37) Product Images from "Parthenolide and DMAPT induce cell death in primitive CML cells through reactive oxygen species, et al. Parthenolide and DMAPT induce cell death in primitive CML cells through reactive oxygen species"

    Article Title: Parthenolide and DMAPT induce cell death in primitive CML cells through reactive oxygen species, et al. Parthenolide and DMAPT induce cell death in primitive CML cells through reactive oxygen species

    Journal: Journal of Cellular and Molecular Medicine

    doi: 10.1111/jcmm.13755

    PTL and DMAPT treatment correlates with NF ‐κB inhibition in CML cells. Leukaemic cell lines and primary CD 34 + lin − cells from CML patients and normal bone marrows were cultured in the absence or presence of 7.5 and 10 μmol/L PTL or DMAPT . Results in (A) represent an immunofluorescence analysis for total p65 and nuclear stain with DAPI in K562 cell line. B, correspond to a representative Western Blot for phospho p65 and p65 total, using LPS as a positive control of NF ‐κB induction and beta‐actin as a loading control. C, Represents analysis of NF ‐κB transcription targets ( NFKB 1 and PLAU ) in cells treated with PTL or DMAPT for 6 h. Scale bar: 10 μm
    Figure Legend Snippet: PTL and DMAPT treatment correlates with NF ‐κB inhibition in CML cells. Leukaemic cell lines and primary CD 34 + lin − cells from CML patients and normal bone marrows were cultured in the absence or presence of 7.5 and 10 μmol/L PTL or DMAPT . Results in (A) represent an immunofluorescence analysis for total p65 and nuclear stain with DAPI in K562 cell line. B, correspond to a representative Western Blot for phospho p65 and p65 total, using LPS as a positive control of NF ‐κB induction and beta‐actin as a loading control. C, Represents analysis of NF ‐κB transcription targets ( NFKB 1 and PLAU ) in cells treated with PTL or DMAPT for 6 h. Scale bar: 10 μm

    Techniques Used: Inhibition, Cell Culture, Immunofluorescence, Staining, Western Blot, Positive Control

    38) Product Images from "Regulation of the T-box transcription factor Tbx3 by the tumour suppressor microRNA-206 in breast cancer"

    Article Title: Regulation of the T-box transcription factor Tbx3 by the tumour suppressor microRNA-206 in breast cancer

    Journal: British Journal of Cancer

    doi: 10.1038/bjc.2016.73

    miR-206-mediated downregulation of Tbx3 impacts its targets, inhibits proliferation and promotes apoptosis. ( A ) Western blot analysis of MCF7 and MDA-MB-231 cells transfected with miR scrambled (NC), miR-206 or α miR-206 for 72 h. Beta-actin was used as a protein-loading control. ( B ) Western blot analysis of ZR75-1 and MDA-MB-231 cells transfected with miR scrambled control (NC) or siRNA-Tbx3 (si-Tbx3). Beta-actin was used as a loading control. ( C ) Bar plots showing per cent colony formation of MDA-MB-231, ZR75-1 and MCF7 cells transfected with miR scrambled control (NC), miR-206, α miR-206, siRNA-Tbx3 (si-Tbx3) or left untreated (UT). Per cent colony formation was determined with untreated set at 100%. ( D ) Bar plots showing relative DNA fragmentation in MDA-MB-231, ZR75-1 and MCF7 cells transfected with miR scrambled control (NC), miR-206, α miR-206, siRNA-Tbx3 (si-Tbx3) or left untreated (UT). Untreated (UT) cells and miR scrambled control (NC) were used as controls. The results are representative of three independent experiment performed in triplicates. Means with error bars representing±s.e.m. * P
    Figure Legend Snippet: miR-206-mediated downregulation of Tbx3 impacts its targets, inhibits proliferation and promotes apoptosis. ( A ) Western blot analysis of MCF7 and MDA-MB-231 cells transfected with miR scrambled (NC), miR-206 or α miR-206 for 72 h. Beta-actin was used as a protein-loading control. ( B ) Western blot analysis of ZR75-1 and MDA-MB-231 cells transfected with miR scrambled control (NC) or siRNA-Tbx3 (si-Tbx3). Beta-actin was used as a loading control. ( C ) Bar plots showing per cent colony formation of MDA-MB-231, ZR75-1 and MCF7 cells transfected with miR scrambled control (NC), miR-206, α miR-206, siRNA-Tbx3 (si-Tbx3) or left untreated (UT). Per cent colony formation was determined with untreated set at 100%. ( D ) Bar plots showing relative DNA fragmentation in MDA-MB-231, ZR75-1 and MCF7 cells transfected with miR scrambled control (NC), miR-206, α miR-206, siRNA-Tbx3 (si-Tbx3) or left untreated (UT). Untreated (UT) cells and miR scrambled control (NC) were used as controls. The results are representative of three independent experiment performed in triplicates. Means with error bars representing±s.e.m. * P

    Techniques Used: Western Blot, Multiple Displacement Amplification, Transfection

    miR-206 expression and Tbx3 knock-down inhibits the 3D growth of breast cancer cells in Matrigel. Tbx3 lacking the 3′ UTR is not repressed by miR-206. ( A ) Phase-contrast images of MDA-MB-231 cells in 3D culture treated with miR-206, α miR-206 or siRNA-Tbx3 (si-Tbx3). Untreated (UT) and miR scrambled control (NC) treated cells were used as controls. ( B ) Bar plots represent the quantification of the number of invasive colonies, mean colony area and number of branching stellate cells. ( C ) Western blot analysis of MDA-MB-231 cells co-transfected with miR scrambled control with either empty vector (NC+V) or with flag-tagged TBX3 expression vector (NC+TBX3), and with miR-206 with empty vector (miR-206+V) or with flag-tagged TBX3 expression vector (miR-206+TBX3). Beta-actin was used as a loading control. ( D ) Phase-contrast images of MDA-MB-231 cells in 3D cultures and co-transfected with miR scrambled control (NC) or miR-206 with either empty vector (V) control or TBX3 expression vector (TBX3). ( E ) Bar plots showing the reduction in invasive phenotype of MDA-MB-231 cells as quantified by counting invasive colonies, mean colony area and the number of branching stellate cells. The results are representatives of three independent experiments. Scale bar: × 4 images, 200 μ m; × 10 images, 50 μ m. Means with error bars representing±s.e.m. * P
    Figure Legend Snippet: miR-206 expression and Tbx3 knock-down inhibits the 3D growth of breast cancer cells in Matrigel. Tbx3 lacking the 3′ UTR is not repressed by miR-206. ( A ) Phase-contrast images of MDA-MB-231 cells in 3D culture treated with miR-206, α miR-206 or siRNA-Tbx3 (si-Tbx3). Untreated (UT) and miR scrambled control (NC) treated cells were used as controls. ( B ) Bar plots represent the quantification of the number of invasive colonies, mean colony area and number of branching stellate cells. ( C ) Western blot analysis of MDA-MB-231 cells co-transfected with miR scrambled control with either empty vector (NC+V) or with flag-tagged TBX3 expression vector (NC+TBX3), and with miR-206 with empty vector (miR-206+V) or with flag-tagged TBX3 expression vector (miR-206+TBX3). Beta-actin was used as a loading control. ( D ) Phase-contrast images of MDA-MB-231 cells in 3D cultures and co-transfected with miR scrambled control (NC) or miR-206 with either empty vector (V) control or TBX3 expression vector (TBX3). ( E ) Bar plots showing the reduction in invasive phenotype of MDA-MB-231 cells as quantified by counting invasive colonies, mean colony area and the number of branching stellate cells. The results are representatives of three independent experiments. Scale bar: × 4 images, 200 μ m; × 10 images, 50 μ m. Means with error bars representing±s.e.m. * P

    Techniques Used: Expressing, Multiple Displacement Amplification, Western Blot, Transfection, Plasmid Preparation

    miR-206 exerts a tumour suppressive function and inhibits tumorsphere formation of breast cancer cells. Phase-contrast × 4 images of tumorspheres of ( A ) MCF7 and ( B ) MDA-MB-231 cells reverse-transfected with miR scrambled control (NC), miR-206, α miR-206, siRNA-Tbx3 (si-Tbx3), miR-206 plus empty vector (miR-206+V) or miR-206 plus TBX3 expression vector (miR-206+TBX3). Bar plots showing the relative sphere number of ( B ) MCF7 and ( E ) MDA-MB-231 cells corresponding to A and B , respectively. Relative sphere number was normalised to miR scrambled control (NC). Images are representative of spheres under the mentioned treatments at the secondary sphere stage. Western blot analysis of ( C ) MCF7 and ( F ) MDA-MB-231 cells co-transfected with miR scrambled control (NC), miR-206, α miR-206, siRNA-Tbx3 (si-Tbx3), miR-206 plus empty vector (miR-206+V) or miR-206 plus TBX3 expression vector (miR-206+TBX3). Beta-actin was used a loading control. Scale bar: 200 μ m. Means with error bars representing±s.e.m. * P
    Figure Legend Snippet: miR-206 exerts a tumour suppressive function and inhibits tumorsphere formation of breast cancer cells. Phase-contrast × 4 images of tumorspheres of ( A ) MCF7 and ( B ) MDA-MB-231 cells reverse-transfected with miR scrambled control (NC), miR-206, α miR-206, siRNA-Tbx3 (si-Tbx3), miR-206 plus empty vector (miR-206+V) or miR-206 plus TBX3 expression vector (miR-206+TBX3). Bar plots showing the relative sphere number of ( B ) MCF7 and ( E ) MDA-MB-231 cells corresponding to A and B , respectively. Relative sphere number was normalised to miR scrambled control (NC). Images are representative of spheres under the mentioned treatments at the secondary sphere stage. Western blot analysis of ( C ) MCF7 and ( F ) MDA-MB-231 cells co-transfected with miR scrambled control (NC), miR-206, α miR-206, siRNA-Tbx3 (si-Tbx3), miR-206 plus empty vector (miR-206+V) or miR-206 plus TBX3 expression vector (miR-206+TBX3). Beta-actin was used a loading control. Scale bar: 200 μ m. Means with error bars representing±s.e.m. * P

    Techniques Used: Multiple Displacement Amplification, Transfection, Plasmid Preparation, Expressing, Western Blot

    Tbx3 is a direct target of miR-206 in breast cancer cells, and restoration of miR-206 decreases Tbx3 mRNA and protein expression in breast cancer cells. ( A ) Putative miR-206 target site within the human Tbx3 3′ UTR as predicted by TargetScan and PicTar algorithms. ( B ) The miR-206 wild-type binding sequence or its mutated form was inserted into the C terminus of the luciferase gene to generate pMIR-TBX3-3′ UTR or pMIR-TBX3-mut-3′ UTR, respectively. ( C ) Bar plots showing relative luciferase activity in HEK-293T cells co-transfected with a luciferase reporter construct containing a fragment of the human Tbx3 3′ UTR encompassing the miR-206 binding site (Tbx3 3′ UTR) or empty vector (V), along with miR scrambled control (NC) or miR-206 for 48 h. Relative luciferase units were normalised to control β -gal reporter vector for transfection efficiency. ( D ) Sequence alignment showing the evolutionary conservation of the 3′ UTR of Tbx3 target site across different species. ( E ) Bar plots showing relative Tbx3 mRNA expression levels, in ZR75-1, MCF7 and MDA-MB-231 cells transfected with miR scrambled control (NC) or miR-206. Analysis for the Tbx3 message levels was performed by the comparative Ct method. Relative abundance was determined from the Ct values using the 2 −ΔΔCt method after normalisation to GAPDH. Results are representative of at least three independent experiments. ( F ) Western blot analysis of MDA-MB-231 and MCF7 cells transfected with miR scrambled control (NC), miR-206 or α miR-206 for 72 h. Beta-actin served as a loading control. Results are representative of three to five independent experiments. Means with error bars representing±s.e.m. ** P
    Figure Legend Snippet: Tbx3 is a direct target of miR-206 in breast cancer cells, and restoration of miR-206 decreases Tbx3 mRNA and protein expression in breast cancer cells. ( A ) Putative miR-206 target site within the human Tbx3 3′ UTR as predicted by TargetScan and PicTar algorithms. ( B ) The miR-206 wild-type binding sequence or its mutated form was inserted into the C terminus of the luciferase gene to generate pMIR-TBX3-3′ UTR or pMIR-TBX3-mut-3′ UTR, respectively. ( C ) Bar plots showing relative luciferase activity in HEK-293T cells co-transfected with a luciferase reporter construct containing a fragment of the human Tbx3 3′ UTR encompassing the miR-206 binding site (Tbx3 3′ UTR) or empty vector (V), along with miR scrambled control (NC) or miR-206 for 48 h. Relative luciferase units were normalised to control β -gal reporter vector for transfection efficiency. ( D ) Sequence alignment showing the evolutionary conservation of the 3′ UTR of Tbx3 target site across different species. ( E ) Bar plots showing relative Tbx3 mRNA expression levels, in ZR75-1, MCF7 and MDA-MB-231 cells transfected with miR scrambled control (NC) or miR-206. Analysis for the Tbx3 message levels was performed by the comparative Ct method. Relative abundance was determined from the Ct values using the 2 −ΔΔCt method after normalisation to GAPDH. Results are representative of at least three independent experiments. ( F ) Western blot analysis of MDA-MB-231 and MCF7 cells transfected with miR scrambled control (NC), miR-206 or α miR-206 for 72 h. Beta-actin served as a loading control. Results are representative of three to five independent experiments. Means with error bars representing±s.e.m. ** P

    Techniques Used: Expressing, Binding Assay, Sequencing, Luciferase, Activity Assay, Transfection, Construct, Plasmid Preparation, Multiple Displacement Amplification, Western Blot

    39) Product Images from "MeCP2 Dependent Heterochromatin Reorganization during Neural Differentiation of a Novel Mecp2-Deficient Embryonic Stem Cell Reporter Line"

    Article Title: MeCP2 Dependent Heterochromatin Reorganization during Neural Differentiation of a Novel Mecp2-Deficient Embryonic Stem Cell Reporter Line

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0047848

    MeCP2 becomes detectable in heterochromatic chromocenters at late differentiation stages. A. Mecp2 wt and Mecp2 −/y tEG cells were differentiated, fixed, and immunostained with antibodies to MeCP2. Tau promoter driven EGFP expression highlights neuronal cells. Additional markers as beta tubulin III (neuronal cell) and GFAP (astroglia) allow discrimination of different cell populations. In immunofluorescence experiments MeCP2 protein is detected as of day 13 (left) and remains constant thereafter (middle) in EGFP + cells. Line profiles across chromocenters highlight MeCP2 protein accumulation at these structures (black line) as well as their intense DAPI signal (blue line). Some GFAP + astroglia revealed a relative weaker MeCP2 signal (black line) compared to neurons. Mecp2 deficient cells (right) were used as control. Bar: 10 µm. B. Immunoblot experiments demonstrate the increase of MeCP2 protein over time in wild type cells (wt) while Mecp2 deficient cells (−/y) show no signal. Beta actin was used as a control for equal loading.
    Figure Legend Snippet: MeCP2 becomes detectable in heterochromatic chromocenters at late differentiation stages. A. Mecp2 wt and Mecp2 −/y tEG cells were differentiated, fixed, and immunostained with antibodies to MeCP2. Tau promoter driven EGFP expression highlights neuronal cells. Additional markers as beta tubulin III (neuronal cell) and GFAP (astroglia) allow discrimination of different cell populations. In immunofluorescence experiments MeCP2 protein is detected as of day 13 (left) and remains constant thereafter (middle) in EGFP + cells. Line profiles across chromocenters highlight MeCP2 protein accumulation at these structures (black line) as well as their intense DAPI signal (blue line). Some GFAP + astroglia revealed a relative weaker MeCP2 signal (black line) compared to neurons. Mecp2 deficient cells (right) were used as control. Bar: 10 µm. B. Immunoblot experiments demonstrate the increase of MeCP2 protein over time in wild type cells (wt) while Mecp2 deficient cells (−/y) show no signal. Beta actin was used as a control for equal loading.

    Techniques Used: Expressing, Immunofluorescence

    40) Product Images from "Spatiotemporal expression and inhibition of prolyl oligopeptidase contradict its involvement in key pathologic mechanisms of kainic acid–induced temporal lobe epilepsy in rats, et al. Spatiotemporal expression and inhibition of prolyl oligopeptidase contradict its involvement in key pathologic mechanisms of kainic acid–induced temporal lobe epilepsy in rats"

    Article Title: Spatiotemporal expression and inhibition of prolyl oligopeptidase contradict its involvement in key pathologic mechanisms of kainic acid–induced temporal lobe epilepsy in rats, et al. Spatiotemporal expression and inhibition of prolyl oligopeptidase contradict its involvement in key pathologic mechanisms of kainic acid–induced temporal lobe epilepsy in rats

    Journal: Epilepsia Open

    doi: 10.1002/epi4.12293

    Prolyl oligopeptidase ( PREP ) specific activity decreased but the expression of PREP remained the same in KYP ‐2047‐treated KASE rats when compared to saline‐treated KASE rats. PREP activity measured in brain lysates at (A) 2‐week and (B) 12‐week time points using the substrate Z‐Gly‐Pro‐ pNA . C, PREP activity measured in plasma using the fluorogenic substrate Z‐Gly‐Pro‐ AMC after 2 weeks of KYP ‐2047 administration. D, Representative western blot of PREP in brain lysates at 2 and 12 weeks after KASE induction. Beta‐actin ( ACTB ) was used as a loading control. Quantification of PREP expression in brain lysates at (E) 2 weeks and (F) 12 weeks after KASE induction. Data are represented as median ± interquartile ranges (* P
    Figure Legend Snippet: Prolyl oligopeptidase ( PREP ) specific activity decreased but the expression of PREP remained the same in KYP ‐2047‐treated KASE rats when compared to saline‐treated KASE rats. PREP activity measured in brain lysates at (A) 2‐week and (B) 12‐week time points using the substrate Z‐Gly‐Pro‐ pNA . C, PREP activity measured in plasma using the fluorogenic substrate Z‐Gly‐Pro‐ AMC after 2 weeks of KYP ‐2047 administration. D, Representative western blot of PREP in brain lysates at 2 and 12 weeks after KASE induction. Beta‐actin ( ACTB ) was used as a loading control. Quantification of PREP expression in brain lysates at (E) 2 weeks and (F) 12 weeks after KASE induction. Data are represented as median ± interquartile ranges (* P

    Techniques Used: Activity Assay, Expressing, Western Blot

    Related Articles

    Western Blot:

    Article Title: FAK activation is required for IGF1R-mediated regulation of EMT, migration, and invasion in mesenchymal triple negative breast cancer cells
    Article Snippet: .. ZEB-1 antibody was from Santa Cruz Biotechnology, Inc. Antibodies against β-actin, and vimentin purchased from Sigma (Saint Louis, MO, USA) and pFAK (Tyr397), total FAK, and E-cadherin from BD Biosciences were used for Western blots according to standard protocol. .. Bound primary antibodies were detected with peroxidase-coupled secondary antibodies (Southern BioTech; Birmingham, AL, USA) and developed by enhanced chemiluminescence (Luminata Classico Western HRP substrate; EMD Millipore Corp.; Billerica, MA, USA).

    Incubation:

    Article Title: Wnt signal activation induces midbrain specification through direct binding of the beta-catenin/TCF4 complex to the EN1 promoter in human pluripotent stem cells
    Article Snippet: .. After incubation with 5% non-fat milk in Tris-buffered saline containing 0.1% Tween 20 (TBS-T) for 1 h at room temperature, the membrane was incubated with primary antibodies (mouse anti-β-catenin (Santa Cruz Biotechnology), mouse anti-EN1 (Abcam, Cambridge, UK), and mouse anti-β-actin (Sigma-Aldrich)) for 1 h at room temperature or overnight at 4 °C. .. The membrane was washed three times with TBS-T and then incubated in a 1:3000 dilution of horseradish peroxidase-conjugated anti-mouse secondary antibody for 30 min at room temperature.

    Article Title: SPARCL1 suppresses metastasis in prostate cancer
    Article Snippet: .. Membranes were incubated with chicken polyclonal antibody specific for human SPARCL1 (Abcam, Cambridge, United Kingdom) at a 1:2000 dilution, and then the membranes were re‐probed using mouse monoclonal antibody specific for human anti‐β‐actin (Sigma Chemical Company, Saint Louis, MO, USA). .. The human gene SPARCL1 ORF (RC207583, OriGene Technologies, Inc, Rockville, MD) was subcloned into pBMN‐I‐GFP (Addgene Inc., Cambridge, MA, USA) to obtain a pBMN‐SPARCL1‐I‐GFP plasmid.

    Article Title: Photodynamic therapy inhibits p-glycoprotein mediated multidrug resistance via JNK activation in human hepatocellular carcinoma using the photosensitizer pheophorbide a
    Article Snippet: .. The membrane was blocked with 10% non-fat milk in Tris buffered saline containing Tween-20 (TBS-T) (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, 0.1% Tween-20) and then incubated with primary human antibodies against β-actin (Sigma), Bcl-2, JNK, p-JNK, p-ERK, procaspase-3 (Santa Cruz), caspase-9 (Stressgen), and P-glycoprotein (Merck) in TBS-T. After incubation with the secondary antibody conjugated with horseradish peroxidase, immunodetected proteins were visualized by using an enhanced chemiluminescence assay kit (Amersham Life Science). ..

    Article Title: ORP150/HSP12A Regulates Purkinje Cell Survival: A Role for Endoplasmic Reticulum Stress in Cerebellar Development
    Article Snippet: .. Tissue extracts were prepared in PBS containing NP-40 (1%), and proteins were separated by SDS-PAGE and transferred to polyvinylidene difluoride paper, followed by incubation with either anti-human ORP150 antibody (1 μg/ml) or anti-human β-actin antibody (1000× dilution; Sigma), the latter as an internal control. .. Where indicated, Western blotting using anti-KDEL monoclonal antibody (Stressgen Biotechnologies Co.; 0.2 μg/ml) was used to assess levels of GRP78 (78 kDa glucose-regulated protein; ).

    Chemiluminescence Immunoassay:

    Article Title: Photodynamic therapy inhibits p-glycoprotein mediated multidrug resistance via JNK activation in human hepatocellular carcinoma using the photosensitizer pheophorbide a
    Article Snippet: .. The membrane was blocked with 10% non-fat milk in Tris buffered saline containing Tween-20 (TBS-T) (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, 0.1% Tween-20) and then incubated with primary human antibodies against β-actin (Sigma), Bcl-2, JNK, p-JNK, p-ERK, procaspase-3 (Santa Cruz), caspase-9 (Stressgen), and P-glycoprotein (Merck) in TBS-T. After incubation with the secondary antibody conjugated with horseradish peroxidase, immunodetected proteins were visualized by using an enhanced chemiluminescence assay kit (Amersham Life Science). ..

    SDS Page:

    Article Title: ORP150/HSP12A Regulates Purkinje Cell Survival: A Role for Endoplasmic Reticulum Stress in Cerebellar Development
    Article Snippet: .. Tissue extracts were prepared in PBS containing NP-40 (1%), and proteins were separated by SDS-PAGE and transferred to polyvinylidene difluoride paper, followed by incubation with either anti-human ORP150 antibody (1 μg/ml) or anti-human β-actin antibody (1000× dilution; Sigma), the latter as an internal control. .. Where indicated, Western blotting using anti-KDEL monoclonal antibody (Stressgen Biotechnologies Co.; 0.2 μg/ml) was used to assess levels of GRP78 (78 kDa glucose-regulated protein; ).

    FLAG-tag:

    Article Title: Seven in Absentia Homolog 2 (Siah2) Protein Is a Regulator of NF-E2-related Factor 2 (Nrf2) *Seven in Absentia Homolog 2 (Siah2) Protein Is a Regulator of NF-E2-related Factor 2 (Nrf2) * ♦
    Article Snippet: .. Anti-c-Myc, anti-DYKDDDDK (FLAG) tag, and anti-HA monoclonal antibodies were purchased from Wako; anti-phosphoserine antibody was from BD Biosciences; anti-human ubiquitin antibody was from Enzo Life Sciences (Farmingdale, NY); anti-human Siah2 antibody was from Santa Cruz Biotechnology (Santa Cruz, CA); anti-human β-actin antibody was from Sigma; and horseradish peroxidase-conjugated anti-rabbit and anti-mouse IgG antibodies were from Bio-Rad. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93
    Millipore antibody against dbh
    Anti-dopamine-β-hydroxylase <t>(DBH)-conjugated</t> saporin (DSAP) produces glucocorticoid resistance in the paraventricular nucleus of the hypothalamus (PVH) of high-fat/high-sucrose choice (HFSC)-fed animals. A–G : combined adrenal weights ( A ), thymus weights ( B ), and corticotropin-releasing hormone (CRH) immunoreactivity (ir) in the PVH ( C and D–G ) in mouse IgG-saporin conjugate <t>(MSAP)</t> and DSAP animals given chow or a HFSC diet for 56 days. MGL, mean gray level. H–K : DBH immunoreactivity in the same sections stained for CRH immunoreactivity ( D–G ) in MSAP ( H and J ) and DSAP ( E and G ) animals. Scale bars = 150 μm. dp, PVH dorsal parvicellular part; mpd, PVH dorsal zone of the medial parvicellular part; mpv, PVH ventral zone of the medial parvicellular part; pm, PVH posterior magnocellular part; pv, PVH periventricular part; V3, 3rd ventricle. Values are means ± SE; n = 5 MSAP-chow and DSAP-HFSC and n = 8 DSAP-chow and MSAP-HFSC. ns, Not significant.
    Antibody Against Dbh, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibody against dbh/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    antibody against dbh - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    99
    Millipore anti α sma
    Preischemic administration of stromal vascular fraction (SVF) inhibited TGF-β1-induced epithelia-mesenchymal transition (EMT) and microvascular rarefaction. (A–C): Relative abundance of E-cadherin/GAPDH (A) and <t>α-SMA/GAPDH</t> (B)
    Anti α Sma, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti α sma/product/Millipore
    Average 99 stars, based on 35 article reviews
    Price from $9.99 to $1999.99
    anti α sma - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    99
    Millipore mouse anti human β actin monoclonal antibody
    Effect of MSK1-mediated p65 Ser276 phosphorylation in IL-1β-induced SCF expression. A. Human lung fibroblasts in culture were transiently co-transfected with the pGL3e/SCF firefly luciferase construct and a Renilla luciferase construct (pRL-TK) as an internal control. Cells were pre-incubated for 1 h with a combination of SB202190 (SB; 3.5 µM) and PD98059 (PD; 20 µM) or with H89 (10 µM) and treated with IL-1β (20 U/ml). After 150 min, cells were harvested for luciferase activity measurement. The results are expressed as the level of pGL3e/SCF constructions' promoter-driven firefly luciferase expression after correcting for the transfection efficiency by pRL-TK luciferase measurements and represented as a percentage of control values. B. Fibroblasts were transfected with control and anti-MSK1 siRNA (100 nM), or transfection medium alone (control). After 48 hours, inhibition of MSK1 with siRNA was controlled by Western blotting in the cell lysate, using anti-MSK1, with <t>anti-β-actin</t> antibodies as a deposit control. Cells were treated with IL-1β (20 U/ml). SCF protein levels were assessed in the supernatant 5 hours after treatment by ELISA. C . Fibroblasts were transfected with WT or “kinase-dead” (KD) MSK1 plasmid (1 µg), WT or S276C p65 plasmids or transfection medium alone (control), and treated with IL-1β (20 U/ml). SCF protein levels were assessed by ELISA in the supernatant obtained 5 hours after treatment. Results are expressed as percentages of control values of three independent experiments performed in fibroblasts from three different donors.
    Mouse Anti Human β Actin Monoclonal Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti human β actin monoclonal antibody/product/Millipore
    Average 99 stars, based on 17 article reviews
    Price from $9.99 to $1999.99
    mouse anti human β actin monoclonal antibody - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    Image Search Results


    Anti-dopamine-β-hydroxylase (DBH)-conjugated saporin (DSAP) produces glucocorticoid resistance in the paraventricular nucleus of the hypothalamus (PVH) of high-fat/high-sucrose choice (HFSC)-fed animals. A–G : combined adrenal weights ( A ), thymus weights ( B ), and corticotropin-releasing hormone (CRH) immunoreactivity (ir) in the PVH ( C and D–G ) in mouse IgG-saporin conjugate (MSAP) and DSAP animals given chow or a HFSC diet for 56 days. MGL, mean gray level. H–K : DBH immunoreactivity in the same sections stained for CRH immunoreactivity ( D–G ) in MSAP ( H and J ) and DSAP ( E and G ) animals. Scale bars = 150 μm. dp, PVH dorsal parvicellular part; mpd, PVH dorsal zone of the medial parvicellular part; mpv, PVH ventral zone of the medial parvicellular part; pm, PVH posterior magnocellular part; pv, PVH periventricular part; V3, 3rd ventricle. Values are means ± SE; n = 5 MSAP-chow and DSAP-HFSC and n = 8 DSAP-chow and MSAP-HFSC. ns, Not significant.

    Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

    Article Title: Obesity, Diabetes and Energy Homeostasis: Catecholaminergic projections into an interconnected forebrain network control the sensitivity of male rats to diet-induced obesity

    doi: 10.1152/ajpregu.00423.2017

    Figure Lengend Snippet: Anti-dopamine-β-hydroxylase (DBH)-conjugated saporin (DSAP) produces glucocorticoid resistance in the paraventricular nucleus of the hypothalamus (PVH) of high-fat/high-sucrose choice (HFSC)-fed animals. A–G : combined adrenal weights ( A ), thymus weights ( B ), and corticotropin-releasing hormone (CRH) immunoreactivity (ir) in the PVH ( C and D–G ) in mouse IgG-saporin conjugate (MSAP) and DSAP animals given chow or a HFSC diet for 56 days. MGL, mean gray level. H–K : DBH immunoreactivity in the same sections stained for CRH immunoreactivity ( D–G ) in MSAP ( H and J ) and DSAP ( E and G ) animals. Scale bars = 150 μm. dp, PVH dorsal parvicellular part; mpd, PVH dorsal zone of the medial parvicellular part; mpv, PVH ventral zone of the medial parvicellular part; pm, PVH posterior magnocellular part; pv, PVH periventricular part; V3, 3rd ventricle. Values are means ± SE; n = 5 MSAP-chow and DSAP-HFSC and n = 8 DSAP-chow and MSAP-HFSC. ns, Not significant.

    Article Snippet: Sections containing the hypothalamus and hindbrains of all MSAP- and DSAP-injected animals were stained using an antibody against DBH (1:10,000 dilution, mouse anti-DBH; catalog no. MAB308, Millipore, Temecula, CA).

    Techniques: Staining

    Preischemic administration of stromal vascular fraction (SVF) inhibited TGF-β1-induced epithelia-mesenchymal transition (EMT) and microvascular rarefaction. (A–C): Relative abundance of E-cadherin/GAPDH (A) and α-SMA/GAPDH (B)

    Journal: Stem Cells Translational Medicine

    Article Title: Preischemic Administration of Nonexpanded Adipose Stromal Vascular Fraction Attenuates Acute Renal Ischemia/Reperfusion Injury and Fibrosis

    doi: 10.5966/sctm.2015-0223

    Figure Lengend Snippet: Preischemic administration of stromal vascular fraction (SVF) inhibited TGF-β1-induced epithelia-mesenchymal transition (EMT) and microvascular rarefaction. (A–C): Relative abundance of E-cadherin/GAPDH (A) and α-SMA/GAPDH (B)

    Article Snippet: The membranes were blocked with 10% skimmed milk for 1 hour and incubated at 4°C overnight with anti-α-SMA, anti-E-cadherin, or anti-glyceraldehyde-3-phosphate dehydrogenase (EMD Millipore), followed by incubation with horseradish peroxidase-conjugated anti-mouse IgG (Cell Signaling, Danvers, MA, ).

    Techniques:

    Effect of MSK1-mediated p65 Ser276 phosphorylation in IL-1β-induced SCF expression. A. Human lung fibroblasts in culture were transiently co-transfected with the pGL3e/SCF firefly luciferase construct and a Renilla luciferase construct (pRL-TK) as an internal control. Cells were pre-incubated for 1 h with a combination of SB202190 (SB; 3.5 µM) and PD98059 (PD; 20 µM) or with H89 (10 µM) and treated with IL-1β (20 U/ml). After 150 min, cells were harvested for luciferase activity measurement. The results are expressed as the level of pGL3e/SCF constructions' promoter-driven firefly luciferase expression after correcting for the transfection efficiency by pRL-TK luciferase measurements and represented as a percentage of control values. B. Fibroblasts were transfected with control and anti-MSK1 siRNA (100 nM), or transfection medium alone (control). After 48 hours, inhibition of MSK1 with siRNA was controlled by Western blotting in the cell lysate, using anti-MSK1, with anti-β-actin antibodies as a deposit control. Cells were treated with IL-1β (20 U/ml). SCF protein levels were assessed in the supernatant 5 hours after treatment by ELISA. C . Fibroblasts were transfected with WT or “kinase-dead” (KD) MSK1 plasmid (1 µg), WT or S276C p65 plasmids or transfection medium alone (control), and treated with IL-1β (20 U/ml). SCF protein levels were assessed by ELISA in the supernatant obtained 5 hours after treatment. Results are expressed as percentages of control values of three independent experiments performed in fibroblasts from three different donors.

    Journal: PLoS ONE

    Article Title: Ser276 Phosphorylation of NF-kB p65 by MSK1 Controls SCF Expression in Inflammation

    doi: 10.1371/journal.pone.0004393

    Figure Lengend Snippet: Effect of MSK1-mediated p65 Ser276 phosphorylation in IL-1β-induced SCF expression. A. Human lung fibroblasts in culture were transiently co-transfected with the pGL3e/SCF firefly luciferase construct and a Renilla luciferase construct (pRL-TK) as an internal control. Cells were pre-incubated for 1 h with a combination of SB202190 (SB; 3.5 µM) and PD98059 (PD; 20 µM) or with H89 (10 µM) and treated with IL-1β (20 U/ml). After 150 min, cells were harvested for luciferase activity measurement. The results are expressed as the level of pGL3e/SCF constructions' promoter-driven firefly luciferase expression after correcting for the transfection efficiency by pRL-TK luciferase measurements and represented as a percentage of control values. B. Fibroblasts were transfected with control and anti-MSK1 siRNA (100 nM), or transfection medium alone (control). After 48 hours, inhibition of MSK1 with siRNA was controlled by Western blotting in the cell lysate, using anti-MSK1, with anti-β-actin antibodies as a deposit control. Cells were treated with IL-1β (20 U/ml). SCF protein levels were assessed in the supernatant 5 hours after treatment by ELISA. C . Fibroblasts were transfected with WT or “kinase-dead” (KD) MSK1 plasmid (1 µg), WT or S276C p65 plasmids or transfection medium alone (control), and treated with IL-1β (20 U/ml). SCF protein levels were assessed by ELISA in the supernatant obtained 5 hours after treatment. Results are expressed as percentages of control values of three independent experiments performed in fibroblasts from three different donors.

    Article Snippet: Immunoblotting used the following antibodies: rabbit anti-human IκB-α polyclonal antibody (1/1000, Calbiochem, La Jolla, CA), mouse anti-human phospho- IκB-α monoclonal antibody, (1/1000, Ab-1, Oncogene Research Product, Boston, MA), rabbit anti-human phospho-Ser276 p65 antibody (1/1000, 3037, Cell Signaling Technology, Danvers MA), rabbit anti-human phospho-Ser536 p65 antibody (1/1000, 3031, Cell Signaling Technology), rabbit anti-human p65 polyclonal antibody (1/200, sc-109, Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti-human CBP polyclonal antibody (1/200, sc-369, Santa Cruz Biotechnology), mouse anti-human β-actin monoclonal antibody (1/5000, Ab-1, Oncogene Research Product), goat anti-human MSK1 (1/200, sc-9392, Santa Cruz Biotechnology.

    Techniques: Expressing, Transfection, Luciferase, Construct, Incubation, Activity Assay, Inhibition, Western Blot, Enzyme-linked Immunosorbent Assay, Plasmid Preparation