rabbit polyclonal anti connexin 43  (Cell Signaling Technology Inc)

 
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    Name:
    Connexin 43 Antibody
    Description:
    Connexin 43 Cx43 is a member of the large family of gap junction proteins Connexins assemble as a hexamer and are transported to the plasma membrane to create a hemichannel that can associate with hemichannels on nearby cells to create cell to cell channels Clusters of these channels assemble to make gap junctions Gap junction communication is important in development and regulation of cell growth Phosphorylation of Cx43 is important in regulating assembly and function of gap junctions 1 2 Ser368 of Cx43 is phosphorylated by protein kinase C PKC after activation by phorbol esters which decreases cell to cell communication 3 Src can interact with and phosphorylate Cx43 to alter gap junction communication 4 5
    Catalog Number:
    3512
    Price:
    None
    Applications:
    Western Blot, Immunohistochemistry, Immunofluorescence
    Category:
    Primary Antibodies
    Source:
    Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues of human connexin 43. Antibodies are purified by protein A and peptide affinity chromatography.
    Reactivity:
    Human Mouse Rat Monkey Zebrafish
    Buy from Supplier


    Structured Review

    Cell Signaling Technology Inc rabbit polyclonal anti connexin 43
    Connexin 43 Cx43 is a member of the large family of gap junction proteins Connexins assemble as a hexamer and are transported to the plasma membrane to create a hemichannel that can associate with hemichannels on nearby cells to create cell to cell channels Clusters of these channels assemble to make gap junctions Gap junction communication is important in development and regulation of cell growth Phosphorylation of Cx43 is important in regulating assembly and function of gap junctions 1 2 Ser368 of Cx43 is phosphorylated by protein kinase C PKC after activation by phorbol esters which decreases cell to cell communication 3 Src can interact with and phosphorylate Cx43 to alter gap junction communication 4 5
    https://www.bioz.com/result/rabbit polyclonal anti connexin 43/product/Cell Signaling Technology Inc
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal anti connexin 43 - by Bioz Stars, 2020-08
    97/100 stars

    Images

    Related Articles

    Staining:

    Article Title: Role of connexins in metastatic breast cancer and melanoma brain colonization
    Article Snippet: .. Four-micron tissue sections were stained with a rabbit polyclonal antibody to Cx43 (#3512, Cell Signaling Technology, Inc.), and a mouse monoclonal antibody to GFAP ( , Spring Bioscience) using Discovery Ultra (Ventana Medical Systems). ..

    Incubation:

    Article Title: Gap Junction Dysfunction in the Prefrontal Cortex Induces Depressive-Like Behaviors in Rats
    Article Snippet: .. The membrane was blocked with 3% BSA and incubated with primary antibody overnight at 4 °C (anti-Cx43, CST, Danvers, MA, 1 : 1000; anti- β -actin, Sigma, 1 : 10 000) followed by horseradish peroxidase (HRP)-conjugated secondary antibody (1 : 5000; KPL, Gaithersburg, MD). ..

    Blocking Assay:

    Article Title: Inhibition of Connexin 43 Hemichannels Alleviates Cerebral Ischemia/Reperfusion Injury via the TLR4 Signaling Pathway
    Article Snippet: .. Then block the membrane with 5% fat-free milk at room temperature for 2 h, and incubate with primary antibodies as follows: anti-Tubulin (Cell Signaling), anti-Cx43 (Cell Signaling), anti-TLR4 (Proteintech), anti-MyD88 (Cell Signaling), anti-NF-κB p65 (Proteintech), anti-IL-1β (Bioss), and anti-TNF-α (Bioss) at 4°C overnight. .. Next day, incubate with secondary horse anti-mouse or goat anti-rabbit antibodies conjugated with horseradish peroxidase (Cell Signaling) for 2 h and visualize by an enhanced chemiluminescence system (ECL).

    Article Title: Arrhythmogenic cardiomyopathy: Identification of desmosomal gene variations and desmosomal protein expression in variation carriers
    Article Snippet: .. Following washing three times with PBS and blocking with goat serum at room temperature for 20 min, the tissue sections were subsequently treated with commercially-available rabbit antibodies targeting connexin-43 (Cx43; 3512; 1:100; Cell Signaling Technology, Inc., Danvers, MA, USA), N-cadherins (ab18203; 1:1,000; Abcam, Cambridge, UK), junction plakoglobin (JUP; 2309; 1:50; Cell Signaling Technology, Inc.), plakophilin 2 (PKP2; ab151402; 1:200; Abcam) and desmoglein 2 (DSG2; ab85632; 1:200; Abcam) overnight at 4°C. .. Following washing with PBS three times, tissue sections were treated with anti-rabbit (pv6001) and anti-mouse (pv6002; both Zhongshan Golden Bridge Biotechnology Co., Ltd., Beijing, China) secondary antibodies and visualized with 3,3′-diaminobenzidine (ZLI-9017; Zhongshan Golden Bridge Biotechnology Co., Ltd.) via light microscopy (DMI-600B; Leica Microsystems, Inc., Buffalo Grove, IL, USA).

    Article Title: Elevated connexin 43 expression in arsenite-and cadmium-transformed human bladder cancer cells, tumor transplants and selected high grade human bladder cancers
    Article Snippet: .. After blocking, the membranes were probed with the Cx43 primary antibody (Cell Signaling Technology, Beverly, MA, Cat# 3512) at a dilution of 1:1000 in blocking buffer overnight at 4°C. .. After washing 3 times with TBS-T, the membranes were incubated with the anti-rabbit secondary antibody (1:2000) in blocking buffer for 1 h. The blots were visualized using the Phototope-HRP (horseradish peroxidase) Western blot detection system (Cell Signaling Technology).

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    Cell Signaling Technology Inc commercially available rabbit antibodies targeting connexin 43
    Representative western blotting of intercalated disc proteins and β-tubulin in the right ventricular myocardium from patients with AC and healthy controls. (A) Western blotting of myocardial intercalated disc proteins in the controls, and AC patients with and without DSG2 variation, along with the quantification of the protein expression; (B) western blotting of myocardial intercalated disc proteins in the controls, and AC patients with and without PKP2 variation, along with the quantification of the protein expression. DSG2, desmoglein 2; JUP, junction plakoglobin; Cx43, connexin 43; PKP2, plakophilin 2.
    Commercially Available Rabbit Antibodies Targeting Connexin 43, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/commercially available rabbit antibodies targeting connexin 43/product/Cell Signaling Technology Inc
    Average 97 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    commercially available rabbit antibodies targeting connexin 43 - by Bioz Stars, 2020-08
    97/100 stars
      Buy from Supplier

    90
    Cell Signaling Technology Inc antibody against gja1
    Immunofluorescence images of <t>Gja1,</t> Gjb1 and Gjb2 in IL1-WT and IL1-KO cells after exposure to CNT-1 and CNT-2. IL1-WT and IL1-KO cells were plated on cover slips and were allowed to attach for 24 h before exposure to CNTs or dispersion media alone for 24 h. Cells were stained with the respective antibodies and fluorescent secondary antibodies were used to detect expression. Hoechst staining was used to visualize cell nuclei. a Representative images for Gja1. b Representative images for Gjb1. c Representative images for Gjb2. Arrows indicate interesting areas with hemichannels and gap junction channels when present between cells. For Gjb1 arrows also point to dividing cells having increased expression of Gjb1. Scale bar: 10 μm
    Antibody Against Gja1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibody against gja1/product/Cell Signaling Technology Inc
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    antibody against gja1 - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    Image Search Results


    Representative western blotting of intercalated disc proteins and β-tubulin in the right ventricular myocardium from patients with AC and healthy controls. (A) Western blotting of myocardial intercalated disc proteins in the controls, and AC patients with and without DSG2 variation, along with the quantification of the protein expression; (B) western blotting of myocardial intercalated disc proteins in the controls, and AC patients with and without PKP2 variation, along with the quantification of the protein expression. DSG2, desmoglein 2; JUP, junction plakoglobin; Cx43, connexin 43; PKP2, plakophilin 2.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Arrhythmogenic cardiomyopathy: Identification of desmosomal gene variations and desmosomal protein expression in variation carriers

    doi: 10.3892/etm.2018.5694

    Figure Lengend Snippet: Representative western blotting of intercalated disc proteins and β-tubulin in the right ventricular myocardium from patients with AC and healthy controls. (A) Western blotting of myocardial intercalated disc proteins in the controls, and AC patients with and without DSG2 variation, along with the quantification of the protein expression; (B) western blotting of myocardial intercalated disc proteins in the controls, and AC patients with and without PKP2 variation, along with the quantification of the protein expression. DSG2, desmoglein 2; JUP, junction plakoglobin; Cx43, connexin 43; PKP2, plakophilin 2.

    Article Snippet: Following washing three times with PBS and blocking with goat serum at room temperature for 20 min, the tissue sections were subsequently treated with commercially-available rabbit antibodies targeting connexin-43 (Cx43; 3512; 1:100; Cell Signaling Technology, Inc., Danvers, MA, USA), N-cadherins (ab18203; 1:1,000; Abcam, Cambridge, UK), junction plakoglobin (JUP; 2309; 1:50; Cell Signaling Technology, Inc.), plakophilin 2 (PKP2; ab151402; 1:200; Abcam) and desmoglein 2 (DSG2; ab85632; 1:200; Abcam) overnight at 4°C.

    Techniques: Western Blot, Expressing

    Immunohistochemistry of the right ventricular myocardium from desmosomal gene variation carriers. (A) Immunostaining of intercalated disc protein in patients with DSG2 L797Q (magnification, ×400); (B) immunostaining of intercalated disc protein in patients with PKP2 S249T and G808fsX30 (magnification, ×400). Scale bar, 50 µm. DSG2, desmoglein 2; N-Cad, N-cadherin; JUP, junction plakoglobin; Cx43, connexin 43; PKP2, plakophilin 2.

    Journal: Experimental and Therapeutic Medicine

    Article Title: Arrhythmogenic cardiomyopathy: Identification of desmosomal gene variations and desmosomal protein expression in variation carriers

    doi: 10.3892/etm.2018.5694

    Figure Lengend Snippet: Immunohistochemistry of the right ventricular myocardium from desmosomal gene variation carriers. (A) Immunostaining of intercalated disc protein in patients with DSG2 L797Q (magnification, ×400); (B) immunostaining of intercalated disc protein in patients with PKP2 S249T and G808fsX30 (magnification, ×400). Scale bar, 50 µm. DSG2, desmoglein 2; N-Cad, N-cadherin; JUP, junction plakoglobin; Cx43, connexin 43; PKP2, plakophilin 2.

    Article Snippet: Following washing three times with PBS and blocking with goat serum at room temperature for 20 min, the tissue sections were subsequently treated with commercially-available rabbit antibodies targeting connexin-43 (Cx43; 3512; 1:100; Cell Signaling Technology, Inc., Danvers, MA, USA), N-cadherins (ab18203; 1:1,000; Abcam, Cambridge, UK), junction plakoglobin (JUP; 2309; 1:50; Cell Signaling Technology, Inc.), plakophilin 2 (PKP2; ab151402; 1:200; Abcam) and desmoglein 2 (DSG2; ab85632; 1:200; Abcam) overnight at 4°C.

    Techniques: Immunohistochemistry, Immunostaining

    The levels of connexin 43 (Cx43) and connexin 45 (Cx45) proteins. Western blotting analysis was performed to examine the levels of Cx43 and Cx45 in left ventricular tissue among the five experimental groups at 4 weeks after coronary artery occlusion surgery. (A) Control group, (B) Model group, (C) Metoprolol group, (D) Low dose WXKL group, and (E) High dose WXKL group. Values are expressed as the mean ± SD. ∗ P

    Journal: BioMed Research International

    Article Title: Effect of Wenxin Granules on Gap Junction and MiR-1 in Rats with Myocardial Infarction

    doi: 10.1155/2017/3495021

    Figure Lengend Snippet: The levels of connexin 43 (Cx43) and connexin 45 (Cx45) proteins. Western blotting analysis was performed to examine the levels of Cx43 and Cx45 in left ventricular tissue among the five experimental groups at 4 weeks after coronary artery occlusion surgery. (A) Control group, (B) Model group, (C) Metoprolol group, (D) Low dose WXKL group, and (E) High dose WXKL group. Values are expressed as the mean ± SD. ∗ P

    Article Snippet: The following antibodies were used: Connexin 43 rabbit antibody (Cell Signaling Technology Inc, USA, #3512), anti-Connexin 45/GJA7 antibody - C-terminal (Abcam, ab135474), PKC delta rabbit antibody (Cell Signaling Technology Inc, #2058), p44/42 MAPK (Erk1/2) rabbit antibody (Cell Signaling Technology Inc, #9102), Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) antibody (Cell Signaling Technology Inc, #9101), Elk-1 rabbit antibody (Cell Signaling Technology Inc, #9182), Phospho-Elk-1 (Ser383) antibody (Cell Signaling Technology Inc, #9181), and SRF (H-300) rabbit antibody (Santa Cruz Biotechnology Inc, USA, sc-13029).

    Techniques: Western Blot

    Results from in-cell Western analyses showing effects of kinase inhibition on total Cx43 expression in rat primary VSMCs following incubation in the presence of (A) 8Br-cAMP (100 μM), (B) 8Br-cGMP (100 μM), (C) BAY (100 nM), and (D) DADS (50 μM) for 270 min . (A) Inhibition of PKA (by PKI), PKG (by DT2), or PKC (by CalC) each independently subdued 8Br-cAMP-induced increase in Cx43 expression. (B,C) Only PKG and PKC inhibition reversed the effects of 8Br-cGMP and BAY, respectively. (D) Stimulation of Cx43 expression in the presence of DADS was not dependent on PKA, PKG, or PKC. (E) Treatment with any individual kinase inhibitor did not affect total basal Cx43 expression compared with vehicle controls. Data are mean ± SE from n = 4–7 for each treatment. Student–Newman–Keuls method for multiple comparisons following one-way ANOVA was used. * p

    Journal: Frontiers in Physiology

    Article Title: Control of Vascular Smooth Muscle Cell Growth by Connexin 43

    doi: 10.3389/fphys.2012.00220

    Figure Lengend Snippet: Results from in-cell Western analyses showing effects of kinase inhibition on total Cx43 expression in rat primary VSMCs following incubation in the presence of (A) 8Br-cAMP (100 μM), (B) 8Br-cGMP (100 μM), (C) BAY (100 nM), and (D) DADS (50 μM) for 270 min . (A) Inhibition of PKA (by PKI), PKG (by DT2), or PKC (by CalC) each independently subdued 8Br-cAMP-induced increase in Cx43 expression. (B,C) Only PKG and PKC inhibition reversed the effects of 8Br-cGMP and BAY, respectively. (D) Stimulation of Cx43 expression in the presence of DADS was not dependent on PKA, PKG, or PKC. (E) Treatment with any individual kinase inhibitor did not affect total basal Cx43 expression compared with vehicle controls. Data are mean ± SE from n = 4–7 for each treatment. Student–Newman–Keuls method for multiple comparisons following one-way ANOVA was used. * p

    Article Snippet: Fixed cells were permeabilized with 0.1% Triton, blocked, and incubated with IR-tagged rabbit antibodies against Cx43 or the phosphorylated forms of Cx43 at Serines 255, 262, 279, and 368 (Cell Signaling) along with mouse α-tubulin (Sigma) as the housekeeping control at 1:500 dilutions overnight at 4°C.

    Techniques: In-Cell ELISA, Inhibition, Expressing, Incubation

    In-cell Western analyses of site-specific Cx43 phosphorylation in rat primary VSMCs in the presence of (A) 8Br-cAMP and (B) DADS . (A) In the presence of 8Br-cAMP, phosphorylation of the MAPK-sensitive Ser279 was significantly increased compared to vehicle controls. (B) The presence of DADS significantly increased phosphorylation at the MAPK sensitive Ser255 and Ser279 sites, p34 cdc2 kinase-sensitive Ser262, and the PKC-sensitive Ser368. Data are mean ± SE from three independent experiments with an n = 4 for Ser279 phosphorylation, and n = 3–4 for Ser255, 262, and 368 phosphorylation. Student–Newman–Keuls method for multiple comparisons following one-way ANOVA was used. * p

    Journal: Frontiers in Physiology

    Article Title: Control of Vascular Smooth Muscle Cell Growth by Connexin 43

    doi: 10.3389/fphys.2012.00220

    Figure Lengend Snippet: In-cell Western analyses of site-specific Cx43 phosphorylation in rat primary VSMCs in the presence of (A) 8Br-cAMP and (B) DADS . (A) In the presence of 8Br-cAMP, phosphorylation of the MAPK-sensitive Ser279 was significantly increased compared to vehicle controls. (B) The presence of DADS significantly increased phosphorylation at the MAPK sensitive Ser255 and Ser279 sites, p34 cdc2 kinase-sensitive Ser262, and the PKC-sensitive Ser368. Data are mean ± SE from three independent experiments with an n = 4 for Ser279 phosphorylation, and n = 3–4 for Ser255, 262, and 368 phosphorylation. Student–Newman–Keuls method for multiple comparisons following one-way ANOVA was used. * p

    Article Snippet: Fixed cells were permeabilized with 0.1% Triton, blocked, and incubated with IR-tagged rabbit antibodies against Cx43 or the phosphorylated forms of Cx43 at Serines 255, 262, 279, and 368 (Cell Signaling) along with mouse α-tubulin (Sigma) as the housekeeping control at 1:500 dilutions overnight at 4°C.

    Techniques: In-Cell ELISA

    Effects of cyclic nucleotide or Cx43 stimulation on VSMC DNA synthesis . In rat primary VSMCs in the presence of 10% serum for 6 h, DADS (50 μM) significantly reduced the rate of DNA synthesis by 20% compared with vehicle controls estimated by BrdU incorporation. Data are mean ± SE from three independent experiments each with an n = 6. Student–Newman–Keuls method for multiple comparisons following one-way ANOVA was used. * p

    Journal: Frontiers in Physiology

    Article Title: Control of Vascular Smooth Muscle Cell Growth by Connexin 43

    doi: 10.3389/fphys.2012.00220

    Figure Lengend Snippet: Effects of cyclic nucleotide or Cx43 stimulation on VSMC DNA synthesis . In rat primary VSMCs in the presence of 10% serum for 6 h, DADS (50 μM) significantly reduced the rate of DNA synthesis by 20% compared with vehicle controls estimated by BrdU incorporation. Data are mean ± SE from three independent experiments each with an n = 6. Student–Newman–Keuls method for multiple comparisons following one-way ANOVA was used. * p

    Article Snippet: Fixed cells were permeabilized with 0.1% Triton, blocked, and incubated with IR-tagged rabbit antibodies against Cx43 or the phosphorylated forms of Cx43 at Serines 255, 262, 279, and 368 (Cell Signaling) along with mouse α-tubulin (Sigma) as the housekeeping control at 1:500 dilutions overnight at 4°C.

    Techniques: DNA Synthesis, BrdU Incorporation Assay

    Total Cx43 expression in VSMCs . (A) In-cell Western blotting for total Cx43 expression in rat primary VSMC homogenates. Incubation of rat primary VSMCs for 270 min in the presence of 8Br-cAMP (100 μM), 8Br-cGMP (100 μM), BAY (100 nM), or DADS (50 μM) significantly stimulated total Cx43 expression normalized to α-tubulin. Data are from two independent experiments each performed in triplicate. (B) After 24-h incubation the stimulation of Cx43 was significantly increased only after DADS treatment with no observable changes for the 8Br-cAMP, 8Br-cGMP, or BAY treatment groups compared with vehicle controls. For these experiments n = 6 for each treatment. Student–Newman–Keuls method for multiple comparisons following one-way ANOVA was used. * p

    Journal: Frontiers in Physiology

    Article Title: Control of Vascular Smooth Muscle Cell Growth by Connexin 43

    doi: 10.3389/fphys.2012.00220

    Figure Lengend Snippet: Total Cx43 expression in VSMCs . (A) In-cell Western blotting for total Cx43 expression in rat primary VSMC homogenates. Incubation of rat primary VSMCs for 270 min in the presence of 8Br-cAMP (100 μM), 8Br-cGMP (100 μM), BAY (100 nM), or DADS (50 μM) significantly stimulated total Cx43 expression normalized to α-tubulin. Data are from two independent experiments each performed in triplicate. (B) After 24-h incubation the stimulation of Cx43 was significantly increased only after DADS treatment with no observable changes for the 8Br-cAMP, 8Br-cGMP, or BAY treatment groups compared with vehicle controls. For these experiments n = 6 for each treatment. Student–Newman–Keuls method for multiple comparisons following one-way ANOVA was used. * p

    Article Snippet: Fixed cells were permeabilized with 0.1% Triton, blocked, and incubated with IR-tagged rabbit antibodies against Cx43 or the phosphorylated forms of Cx43 at Serines 255, 262, 279, and 368 (Cell Signaling) along with mouse α-tubulin (Sigma) as the housekeeping control at 1:500 dilutions overnight at 4°C.

    Techniques: Expressing, In-Cell ELISA, Incubation

    Effects of cyclic nucleotide or Cx43 stimulation on VSMC proliferation . (A) In rat primary VSMCs in the presence of 10% serum for 72 h and using the MTT assay, 8Br-cAMP (100 μM), the sGC stimulator BAY (100 nM) or the garlic-derived Cx inducer DADS (50 μM) significantly reduced rat primary VSMC numbers compared with vehicle controls. Data are mean ± SEM with an n = 6. (B) Using hemocytometry after 72 h, both 8Br-cAMP and BAY significantly reduced cell numbers by ∼40%, and the presence of DADS reduced cell numbers by ∼20% ( p = 0.06). Data are mean ± SEM of two independent experiments each with n = 3–4. (C) MTT assay performed on VSMCs after 12 h shows no observable changes in any treatment group. Student–Newman–Keuls method for multiple comparisons following one-way ANOVA was used. * p

    Journal: Frontiers in Physiology

    Article Title: Control of Vascular Smooth Muscle Cell Growth by Connexin 43

    doi: 10.3389/fphys.2012.00220

    Figure Lengend Snippet: Effects of cyclic nucleotide or Cx43 stimulation on VSMC proliferation . (A) In rat primary VSMCs in the presence of 10% serum for 72 h and using the MTT assay, 8Br-cAMP (100 μM), the sGC stimulator BAY (100 nM) or the garlic-derived Cx inducer DADS (50 μM) significantly reduced rat primary VSMC numbers compared with vehicle controls. Data are mean ± SEM with an n = 6. (B) Using hemocytometry after 72 h, both 8Br-cAMP and BAY significantly reduced cell numbers by ∼40%, and the presence of DADS reduced cell numbers by ∼20% ( p = 0.06). Data are mean ± SEM of two independent experiments each with n = 3–4. (C) MTT assay performed on VSMCs after 12 h shows no observable changes in any treatment group. Student–Newman–Keuls method for multiple comparisons following one-way ANOVA was used. * p

    Article Snippet: Fixed cells were permeabilized with 0.1% Triton, blocked, and incubated with IR-tagged rabbit antibodies against Cx43 or the phosphorylated forms of Cx43 at Serines 255, 262, 279, and 368 (Cell Signaling) along with mouse α-tubulin (Sigma) as the housekeeping control at 1:500 dilutions overnight at 4°C.

    Techniques: MTT Assay, Derivative Assay

    Immunofluorescence images of Gja1, Gjb1 and Gjb2 in IL1-WT and IL1-KO cells after exposure to CNT-1 and CNT-2. IL1-WT and IL1-KO cells were plated on cover slips and were allowed to attach for 24 h before exposure to CNTs or dispersion media alone for 24 h. Cells were stained with the respective antibodies and fluorescent secondary antibodies were used to detect expression. Hoechst staining was used to visualize cell nuclei. a Representative images for Gja1. b Representative images for Gjb1. c Representative images for Gjb2. Arrows indicate interesting areas with hemichannels and gap junction channels when present between cells. For Gjb1 arrows also point to dividing cells having increased expression of Gjb1. Scale bar: 10 μm

    Journal: Journal of Cell Communication and Signaling

    Article Title: Effects of carbon nanotubes on intercellular communication and involvement of IL-1 genes

    doi: 10.1007/s12079-016-0323-0

    Figure Lengend Snippet: Immunofluorescence images of Gja1, Gjb1 and Gjb2 in IL1-WT and IL1-KO cells after exposure to CNT-1 and CNT-2. IL1-WT and IL1-KO cells were plated on cover slips and were allowed to attach for 24 h before exposure to CNTs or dispersion media alone for 24 h. Cells were stained with the respective antibodies and fluorescent secondary antibodies were used to detect expression. Hoechst staining was used to visualize cell nuclei. a Representative images for Gja1. b Representative images for Gjb1. c Representative images for Gjb2. Arrows indicate interesting areas with hemichannels and gap junction channels when present between cells. For Gjb1 arrows also point to dividing cells having increased expression of Gjb1. Scale bar: 10 μm

    Article Snippet: Intercellular observation of connexin proteins by immunofluorescence (IF) IL1-WT and IL1-KO cells were cultured on cover slips and were allowed to attach for 24 h before exposure to 5 μg/ml of CNT-1 and CNT-2 for 24 h. Cells were then fixed for 20 min in 4 % paraformaldehyde and permeabilized with 0.1 % Triton X-100 in PBS for 5 min. Cover slips were blocked by incubation with 5 % BSA in 0.1 % PBS-Triton X-100 for 1 h at room temperature and incubated overnight with a primary antibody against Gja1 (rabbit connexin 43 polyclonal antibody, Cell Signaling Technology), Gjb1 (mouse anti-connexin-32 monoclonal antibody, Millipore) or Gjb2 (goat anti-Gjb2 polyclonal antibody, Abcam) in 3 % BSA in PBS at 4 °C in a humidified chamber.

    Techniques: Immunofluorescence, Staining, Expressing

    Gja1, Gjb1 and Gjb2 protein levels in IL1-WT or IL1-KO cells after exposure to dispersion media alone, CNT-1 or CNT-2. a Representative western blots for Gja1, Gjb1 and Gjb2 after exposure to CNT-1. b Representative western blots for Gja1, Gjb1 and Gjb2 after exposure to CNT-2. Tubulin was used as a loading control. c Quantification of Gja1, Gjb1 and Gjb2 protein levels normalized to tubulin after exposure to CNT-1. d Quantification after exposure to CNT-2. For the bar graphs the values represent the mean ± standard error (SE) of three independent experiments. * P

    Journal: Journal of Cell Communication and Signaling

    Article Title: Effects of carbon nanotubes on intercellular communication and involvement of IL-1 genes

    doi: 10.1007/s12079-016-0323-0

    Figure Lengend Snippet: Gja1, Gjb1 and Gjb2 protein levels in IL1-WT or IL1-KO cells after exposure to dispersion media alone, CNT-1 or CNT-2. a Representative western blots for Gja1, Gjb1 and Gjb2 after exposure to CNT-1. b Representative western blots for Gja1, Gjb1 and Gjb2 after exposure to CNT-2. Tubulin was used as a loading control. c Quantification of Gja1, Gjb1 and Gjb2 protein levels normalized to tubulin after exposure to CNT-1. d Quantification after exposure to CNT-2. For the bar graphs the values represent the mean ± standard error (SE) of three independent experiments. * P

    Article Snippet: Intercellular observation of connexin proteins by immunofluorescence (IF) IL1-WT and IL1-KO cells were cultured on cover slips and were allowed to attach for 24 h before exposure to 5 μg/ml of CNT-1 and CNT-2 for 24 h. Cells were then fixed for 20 min in 4 % paraformaldehyde and permeabilized with 0.1 % Triton X-100 in PBS for 5 min. Cover slips were blocked by incubation with 5 % BSA in 0.1 % PBS-Triton X-100 for 1 h at room temperature and incubated overnight with a primary antibody against Gja1 (rabbit connexin 43 polyclonal antibody, Cell Signaling Technology), Gjb1 (mouse anti-connexin-32 monoclonal antibody, Millipore) or Gjb2 (goat anti-Gjb2 polyclonal antibody, Abcam) in 3 % BSA in PBS at 4 °C in a humidified chamber.

    Techniques: Western Blot

    Gja1 , Gjb1 and Gjb2 mRNA expression levels in IL1-WT or IL1-KO cells after exposure to dispersion media alone, CNT-1 or CNT-2 . a Gja1 mRNA expression levels investigated by qPCR after exposure to CNT-1 and CNT-2 for 24 h. b Gjb1 mRNA expression levels after exposure to the CNTs for 24 h. c Gjb2 mRNA expression levels after exposure to the CNTs for 24 h. Values represent the mean ± standard error (SE) of three independent experiments performed in triplicate. * P

    Journal: Journal of Cell Communication and Signaling

    Article Title: Effects of carbon nanotubes on intercellular communication and involvement of IL-1 genes

    doi: 10.1007/s12079-016-0323-0

    Figure Lengend Snippet: Gja1 , Gjb1 and Gjb2 mRNA expression levels in IL1-WT or IL1-KO cells after exposure to dispersion media alone, CNT-1 or CNT-2 . a Gja1 mRNA expression levels investigated by qPCR after exposure to CNT-1 and CNT-2 for 24 h. b Gjb1 mRNA expression levels after exposure to the CNTs for 24 h. c Gjb2 mRNA expression levels after exposure to the CNTs for 24 h. Values represent the mean ± standard error (SE) of three independent experiments performed in triplicate. * P

    Article Snippet: Intercellular observation of connexin proteins by immunofluorescence (IF) IL1-WT and IL1-KO cells were cultured on cover slips and were allowed to attach for 24 h before exposure to 5 μg/ml of CNT-1 and CNT-2 for 24 h. Cells were then fixed for 20 min in 4 % paraformaldehyde and permeabilized with 0.1 % Triton X-100 in PBS for 5 min. Cover slips were blocked by incubation with 5 % BSA in 0.1 % PBS-Triton X-100 for 1 h at room temperature and incubated overnight with a primary antibody against Gja1 (rabbit connexin 43 polyclonal antibody, Cell Signaling Technology), Gjb1 (mouse anti-connexin-32 monoclonal antibody, Millipore) or Gjb2 (goat anti-Gjb2 polyclonal antibody, Abcam) in 3 % BSA in PBS at 4 °C in a humidified chamber.

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Localization of Gja1, Gjb1 and Gjb2 protein after CNT exposure. Cytosolic ( c ) and integral membrane/membrane-associated ( m ) proteins were separated into two fractions. Western blot analysis was used to investigate the localization of Gja1, Gjb1 and Gjb2. Antibodies against NaK ATPase α1 and α-Tubulin were used to detect the membrane and cytosolic fractions, respectively. The images shown are representative of two independent experiments

    Journal: Journal of Cell Communication and Signaling

    Article Title: Effects of carbon nanotubes on intercellular communication and involvement of IL-1 genes

    doi: 10.1007/s12079-016-0323-0

    Figure Lengend Snippet: Localization of Gja1, Gjb1 and Gjb2 protein after CNT exposure. Cytosolic ( c ) and integral membrane/membrane-associated ( m ) proteins were separated into two fractions. Western blot analysis was used to investigate the localization of Gja1, Gjb1 and Gjb2. Antibodies against NaK ATPase α1 and α-Tubulin were used to detect the membrane and cytosolic fractions, respectively. The images shown are representative of two independent experiments

    Article Snippet: Intercellular observation of connexin proteins by immunofluorescence (IF) IL1-WT and IL1-KO cells were cultured on cover slips and were allowed to attach for 24 h before exposure to 5 μg/ml of CNT-1 and CNT-2 for 24 h. Cells were then fixed for 20 min in 4 % paraformaldehyde and permeabilized with 0.1 % Triton X-100 in PBS for 5 min. Cover slips were blocked by incubation with 5 % BSA in 0.1 % PBS-Triton X-100 for 1 h at room temperature and incubated overnight with a primary antibody against Gja1 (rabbit connexin 43 polyclonal antibody, Cell Signaling Technology), Gjb1 (mouse anti-connexin-32 monoclonal antibody, Millipore) or Gjb2 (goat anti-Gjb2 polyclonal antibody, Abcam) in 3 % BSA in PBS at 4 °C in a humidified chamber.

    Techniques: Western Blot