connexin antibodies  (Alomone Labs)


Bioz Verified Symbol Alomone Labs is a verified supplier
Bioz Manufacturer Symbol Alomone Labs manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 91

    Structured Review

    Alomone Labs connexin antibodies
    Validation and optimization of <t>Connexin</t> antibodies used in this study. (a–d) Cx37 labeling revealed just a few puncta (green) on blood vessels (blue) in pure PFA fixative (a) and entire outlines of contractile cells in carbodiimide-based fixatives (b and c). Ganglion cells were more visible in PFA fixative (asterisks); the lowest background was in carbodiimide. Pre-absorption of the antibody with a control peptide eliminated the labeling (d). (e–h) Cx40 (green) was associated with outlines of endothelial cells (blue) in all fixative with the less background in carbodiimide (f). Incubation of primary antibody eliminated the labeling (h) with the exception of labeled microglia (arrow). (i–k) Cx43s-positive puncta (green, Sigma antibody) were associated with vasculature in PFA fixative (i); in addition string-like structures along endothelial cell contacts (blue) became visible in carbodiimide fixative (j–k). (l) Omission of the primary antibody eliminated punctate and string-like structures; the microglia was labeled by the secondary antibody (arrow). (m–p) Cx43a produced identical pattern as Cx43s in carbodiimide containing fixatives (n, o). In pure PFA fixative, this antibody labeled well punctate and string-like structures; however the background was significantly higher (m). Incubation of the primary antibodies with control peptide eliminated the specific labeling (p). (q–s) Morphology of DsRed expressing pericytes was best preserved in PFA-containing fixatives (q and s); the DsRed labeling in pericyte processes was lost in pure carbodiimide fixative (q). (t) Summary of fixation procedure. Red crosses indicate signal, black crosses – background; the number of crosses indicates intensity. Scale bar 10 μm. Cx43s – Cx43 antibody from Sigma, Cx43a – Cx43 antibody from Alomone, NB – Neurobiotin.
    Connexin Antibodies, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/connexin antibodies/product/Alomone Labs
    Average 91 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    connexin antibodies - by Bioz Stars, 2022-09
    91/100 stars

    Images

    1) Product Images from "Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina"

    Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

    Journal: The Journal of comparative neurology

    doi: 10.1002/cne.24699

    Validation and optimization of Connexin antibodies used in this study. (a–d) Cx37 labeling revealed just a few puncta (green) on blood vessels (blue) in pure PFA fixative (a) and entire outlines of contractile cells in carbodiimide-based fixatives (b and c). Ganglion cells were more visible in PFA fixative (asterisks); the lowest background was in carbodiimide. Pre-absorption of the antibody with a control peptide eliminated the labeling (d). (e–h) Cx40 (green) was associated with outlines of endothelial cells (blue) in all fixative with the less background in carbodiimide (f). Incubation of primary antibody eliminated the labeling (h) with the exception of labeled microglia (arrow). (i–k) Cx43s-positive puncta (green, Sigma antibody) were associated with vasculature in PFA fixative (i); in addition string-like structures along endothelial cell contacts (blue) became visible in carbodiimide fixative (j–k). (l) Omission of the primary antibody eliminated punctate and string-like structures; the microglia was labeled by the secondary antibody (arrow). (m–p) Cx43a produced identical pattern as Cx43s in carbodiimide containing fixatives (n, o). In pure PFA fixative, this antibody labeled well punctate and string-like structures; however the background was significantly higher (m). Incubation of the primary antibodies with control peptide eliminated the specific labeling (p). (q–s) Morphology of DsRed expressing pericytes was best preserved in PFA-containing fixatives (q and s); the DsRed labeling in pericyte processes was lost in pure carbodiimide fixative (q). (t) Summary of fixation procedure. Red crosses indicate signal, black crosses – background; the number of crosses indicates intensity. Scale bar 10 μm. Cx43s – Cx43 antibody from Sigma, Cx43a – Cx43 antibody from Alomone, NB – Neurobiotin.
    Figure Legend Snippet: Validation and optimization of Connexin antibodies used in this study. (a–d) Cx37 labeling revealed just a few puncta (green) on blood vessels (blue) in pure PFA fixative (a) and entire outlines of contractile cells in carbodiimide-based fixatives (b and c). Ganglion cells were more visible in PFA fixative (asterisks); the lowest background was in carbodiimide. Pre-absorption of the antibody with a control peptide eliminated the labeling (d). (e–h) Cx40 (green) was associated with outlines of endothelial cells (blue) in all fixative with the less background in carbodiimide (f). Incubation of primary antibody eliminated the labeling (h) with the exception of labeled microglia (arrow). (i–k) Cx43s-positive puncta (green, Sigma antibody) were associated with vasculature in PFA fixative (i); in addition string-like structures along endothelial cell contacts (blue) became visible in carbodiimide fixative (j–k). (l) Omission of the primary antibody eliminated punctate and string-like structures; the microglia was labeled by the secondary antibody (arrow). (m–p) Cx43a produced identical pattern as Cx43s in carbodiimide containing fixatives (n, o). In pure PFA fixative, this antibody labeled well punctate and string-like structures; however the background was significantly higher (m). Incubation of the primary antibodies with control peptide eliminated the specific labeling (p). (q–s) Morphology of DsRed expressing pericytes was best preserved in PFA-containing fixatives (q and s); the DsRed labeling in pericyte processes was lost in pure carbodiimide fixative (q). (t) Summary of fixation procedure. Red crosses indicate signal, black crosses – background; the number of crosses indicates intensity. Scale bar 10 μm. Cx43s – Cx43 antibody from Sigma, Cx43a – Cx43 antibody from Alomone, NB – Neurobiotin.

    Techniques Used: Labeling, Incubation, Produced, Expressing

    2) Product Images from "Nicotine protects rat hypoglossal motoneurons from excitotoxic death via downregulation of connexin 36"

    Article Title: Nicotine protects rat hypoglossal motoneurons from excitotoxic death via downregulation of connexin 36

    Journal: Cell Death & Disease

    doi: 10.1038/cddis.2017.232

    Brainstem Cx36 expression decreases because of nicotine or carbenoxolone application. ( a ) Example of western immunoblotting (top) showing the expression of Cx36 in brainstems incubated for 4 h in Krebs, or treated with TBOA, nicotine + TBOA, nicotine, or carbenoxolone + TBOA. Histograms (bottom) quantifying the significantly decreased Cx36 levels after treatment with nicotine + TBOA (n, N =9) or carbenoxolone + TBOA (n, N =5; Kruskal–Wallis one-way ANOVA: P =0.015 among groups). ( b and c ) Example of western blot (top) showing Cx36 expression in membrane ( b ) or the cytoplasmic ( c ) fractions of samples treated as described above. Histograms illustrate how nicotine alone or co-applied induces a significant decrease of Cx36 expression in the membrane pool ( b , bottom; Kruskal–Wallis one-way ANOVA: P =0.069 among groups; Mann–Whitney test: * P =0.016 for sham versus nicotine + TBOA and ** P =0.008 for sham versus nicotine; n, N =5) and an increase in the cytoplasmic one ( c , bottom; Kruskal–Wallis one-way ANOVA: P =0.014 among groups; Tukey test: * P
    Figure Legend Snippet: Brainstem Cx36 expression decreases because of nicotine or carbenoxolone application. ( a ) Example of western immunoblotting (top) showing the expression of Cx36 in brainstems incubated for 4 h in Krebs, or treated with TBOA, nicotine + TBOA, nicotine, or carbenoxolone + TBOA. Histograms (bottom) quantifying the significantly decreased Cx36 levels after treatment with nicotine + TBOA (n, N =9) or carbenoxolone + TBOA (n, N =5; Kruskal–Wallis one-way ANOVA: P =0.015 among groups). ( b and c ) Example of western blot (top) showing Cx36 expression in membrane ( b ) or the cytoplasmic ( c ) fractions of samples treated as described above. Histograms illustrate how nicotine alone or co-applied induces a significant decrease of Cx36 expression in the membrane pool ( b , bottom; Kruskal–Wallis one-way ANOVA: P =0.069 among groups; Mann–Whitney test: * P =0.016 for sham versus nicotine + TBOA and ** P =0.008 for sham versus nicotine; n, N =5) and an increase in the cytoplasmic one ( c , bottom; Kruskal–Wallis one-way ANOVA: P =0.014 among groups; Tukey test: * P

    Techniques Used: Expressing, Western Blot, Incubation, MANN-WHITNEY

    Cx36 mRNA levels and Hsp70 expression. ( a ) Plot illustrating unchanged values of Cx36 mRNA levels among different treatments. Fractional Cx36 expression (AU): 1±0.0, sham; 0.9±0.2, TBOA; 1±0.3, nicotine + TBOA; 1.3±0.2, nicotine (n, N =6). ( b ) Example of Hsp70 immunoblotting. ( c ) Histogram quantifying the significant increase of Hsp70 expression after nicotine + TBOA treatment (Kruskal–Wallis one-way ANOVA: P =0.027 among groups; Holm–Sidak method: * P
    Figure Legend Snippet: Cx36 mRNA levels and Hsp70 expression. ( a ) Plot illustrating unchanged values of Cx36 mRNA levels among different treatments. Fractional Cx36 expression (AU): 1±0.0, sham; 0.9±0.2, TBOA; 1±0.3, nicotine + TBOA; 1.3±0.2, nicotine (n, N =6). ( b ) Example of Hsp70 immunoblotting. ( c ) Histogram quantifying the significant increase of Hsp70 expression after nicotine + TBOA treatment (Kruskal–Wallis one-way ANOVA: P =0.027 among groups; Holm–Sidak method: * P

    Techniques Used: Expressing

    Idealized diagram to account for the toxic effects by TBOA and their antagonism by nicotine on HMs. ( a ) In the presence of TBOA, glutamate (filled red circles) released from presynaptic terminals acts on glutamatergic receptors to depolarize HMs. TBOA-mediated inhibition of glutamate uptake by astrocytes leads to intensification of glutamate effects with build-up of intracellular free Ca 2+ , which acts on mitochondria and favors production of ROS, release of AIF and DNA damage. This process is greatly amplified by the bidirectional (red arrows) operation of gap junctions (Cx36; filled blue cylinders) that spread and recruit HMs into a hyperexcitable state with deleterious consequences on cell survival. ( b ) In the presence of nicotine, the neurotoxic effect of TBOA is largely attenuated. Nicotine exerts multiple effects via nAChRs located on presynaptical terminals (with consequent decrease in glutamate release, and increase in the inhibitory response), HM membrane and even mitochondria. Through these effects, nicotine strongly decreases the motoneuron coupling via gap junctions through redistribution of Cx36 subunits, therefore disjoining motoneurons from their collective excitatory behavior. Our hypothesis is that this process is accompanied by inhibition of Cx36 activity, reduction in production of ROS and facilitation of the effect of Hsp70 to sequester AIF and prevent its nuclear migration. For references, see text
    Figure Legend Snippet: Idealized diagram to account for the toxic effects by TBOA and their antagonism by nicotine on HMs. ( a ) In the presence of TBOA, glutamate (filled red circles) released from presynaptic terminals acts on glutamatergic receptors to depolarize HMs. TBOA-mediated inhibition of glutamate uptake by astrocytes leads to intensification of glutamate effects with build-up of intracellular free Ca 2+ , which acts on mitochondria and favors production of ROS, release of AIF and DNA damage. This process is greatly amplified by the bidirectional (red arrows) operation of gap junctions (Cx36; filled blue cylinders) that spread and recruit HMs into a hyperexcitable state with deleterious consequences on cell survival. ( b ) In the presence of nicotine, the neurotoxic effect of TBOA is largely attenuated. Nicotine exerts multiple effects via nAChRs located on presynaptical terminals (with consequent decrease in glutamate release, and increase in the inhibitory response), HM membrane and even mitochondria. Through these effects, nicotine strongly decreases the motoneuron coupling via gap junctions through redistribution of Cx36 subunits, therefore disjoining motoneurons from their collective excitatory behavior. Our hypothesis is that this process is accompanied by inhibition of Cx36 activity, reduction in production of ROS and facilitation of the effect of Hsp70 to sequester AIF and prevent its nuclear migration. For references, see text

    Techniques Used: Inhibition, Amplification, Activity Assay, Migration

    Nicotine and carbenoxolone induce significant reduction in Cx36 expression. ( a ) Example of HMs after 4-h incubation in Krebs (sham), TBOA, nicotine + TBOA, nicotine, or carbenoxolone + TBOA. HMs were labeled with SMI 32 (green pseudocolor; left column), as motoneuronal marker, or Cx36 (red pseudocolor; middle column). The right column shows merged images where DAPI (blue pseudocolor) is used to visualize nuclei. ( b ) Histograms quantify the Cx36 signal decrease after 4h of nicotine + TBOA (Kruskal–Wallis one-way ANOVA: P =0.005 among groups; Dunn’s method: * P
    Figure Legend Snippet: Nicotine and carbenoxolone induce significant reduction in Cx36 expression. ( a ) Example of HMs after 4-h incubation in Krebs (sham), TBOA, nicotine + TBOA, nicotine, or carbenoxolone + TBOA. HMs were labeled with SMI 32 (green pseudocolor; left column), as motoneuronal marker, or Cx36 (red pseudocolor; middle column). The right column shows merged images where DAPI (blue pseudocolor) is used to visualize nuclei. ( b ) Histograms quantify the Cx36 signal decrease after 4h of nicotine + TBOA (Kruskal–Wallis one-way ANOVA: P =0.005 among groups; Dunn’s method: * P

    Techniques Used: Expressing, Incubation, Labeling, Marker

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 91
    Alomone Labs connexin antibodies
    Validation and optimization of <t>Connexin</t> antibodies used in this study. (a–d) Cx37 labeling revealed just a few puncta (green) on blood vessels (blue) in pure PFA fixative (a) and entire outlines of contractile cells in carbodiimide-based fixatives (b and c). Ganglion cells were more visible in PFA fixative (asterisks); the lowest background was in carbodiimide. Pre-absorption of the antibody with a control peptide eliminated the labeling (d). (e–h) Cx40 (green) was associated with outlines of endothelial cells (blue) in all fixative with the less background in carbodiimide (f). Incubation of primary antibody eliminated the labeling (h) with the exception of labeled microglia (arrow). (i–k) Cx43s-positive puncta (green, Sigma antibody) were associated with vasculature in PFA fixative (i); in addition string-like structures along endothelial cell contacts (blue) became visible in carbodiimide fixative (j–k). (l) Omission of the primary antibody eliminated punctate and string-like structures; the microglia was labeled by the secondary antibody (arrow). (m–p) Cx43a produced identical pattern as Cx43s in carbodiimide containing fixatives (n, o). In pure PFA fixative, this antibody labeled well punctate and string-like structures; however the background was significantly higher (m). Incubation of the primary antibodies with control peptide eliminated the specific labeling (p). (q–s) Morphology of DsRed expressing pericytes was best preserved in PFA-containing fixatives (q and s); the DsRed labeling in pericyte processes was lost in pure carbodiimide fixative (q). (t) Summary of fixation procedure. Red crosses indicate signal, black crosses – background; the number of crosses indicates intensity. Scale bar 10 μm. Cx43s – Cx43 antibody from Sigma, Cx43a – Cx43 antibody from Alomone, NB – Neurobiotin.
    Connexin Antibodies, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/connexin antibodies/product/Alomone Labs
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    connexin antibodies - by Bioz Stars, 2022-09
    91/100 stars
      Buy from Supplier

    91
    Alomone Labs cx26
    Expression profile of CX45, CX36, and <t>CX26</t> transcripts in the human fetal cortex, from 17 to 23 GW. ( A ) Log10 FPKM values derived by the HISAT-StringTie pathway of data analysis. ( B ) Log10 normalized counts derived by the STAR-HTSeq-DESeq2 data analysis pathway. ( C ) Expression profile of 2 predominant transcripts of Cx45 (ENST00000592524.5 [CX45_524.5] and ENST00000426548.5 [Cx45_548.5]) in comparison with Cx36 and Cx26 with HISAT-StringTie pathway of analysis from 17 to 23 GW in the human fetal cortex. In panels A – C , n = 4, 17 GW; n = 2, 18 GW; n = 5, 19 GW; and n = 1, 23 GW. ( D ) Log10 normalized counts of the Cx and pannexin isoforms expressed in the human fetal cortex, age range of 17–23 GW (STAR-HTSeq-DESeq2 data analysis pathway) ( n = 7). ( E ) Differential expression analysis of human fetal samples at 17 and 19 GW, by the STAR-HTSeq-DESeq2 method ( n = 3, 17 GW; n = 2, 19 GW). ( F ) Sex determination based on XIST expression and ratio of X-to-Y coverage in each human subject used in the study ( n = 7). ( G ) Differential expression analysis of male ( n = 4) and female ( n = 3) human fetal samples from 17 to 19 GW, by the STAR-HTSeq-DESeq2 method. ( H ) Differential expression analysis of cortex ( n = 3) and full brain ( n = 3) human fetal samples at 17 GW by the STAR-HTSeq-DESeq2 method. * P adj
    Cx26, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cx26/product/Alomone Labs
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cx26 - by Bioz Stars, 2022-09
    91/100 stars
      Buy from Supplier

    Image Search Results


    Validation and optimization of Connexin antibodies used in this study. (a–d) Cx37 labeling revealed just a few puncta (green) on blood vessels (blue) in pure PFA fixative (a) and entire outlines of contractile cells in carbodiimide-based fixatives (b and c). Ganglion cells were more visible in PFA fixative (asterisks); the lowest background was in carbodiimide. Pre-absorption of the antibody with a control peptide eliminated the labeling (d). (e–h) Cx40 (green) was associated with outlines of endothelial cells (blue) in all fixative with the less background in carbodiimide (f). Incubation of primary antibody eliminated the labeling (h) with the exception of labeled microglia (arrow). (i–k) Cx43s-positive puncta (green, Sigma antibody) were associated with vasculature in PFA fixative (i); in addition string-like structures along endothelial cell contacts (blue) became visible in carbodiimide fixative (j–k). (l) Omission of the primary antibody eliminated punctate and string-like structures; the microglia was labeled by the secondary antibody (arrow). (m–p) Cx43a produced identical pattern as Cx43s in carbodiimide containing fixatives (n, o). In pure PFA fixative, this antibody labeled well punctate and string-like structures; however the background was significantly higher (m). Incubation of the primary antibodies with control peptide eliminated the specific labeling (p). (q–s) Morphology of DsRed expressing pericytes was best preserved in PFA-containing fixatives (q and s); the DsRed labeling in pericyte processes was lost in pure carbodiimide fixative (q). (t) Summary of fixation procedure. Red crosses indicate signal, black crosses – background; the number of crosses indicates intensity. Scale bar 10 μm. Cx43s – Cx43 antibody from Sigma, Cx43a – Cx43 antibody from Alomone, NB – Neurobiotin.

    Journal: The Journal of comparative neurology

    Article Title: Domain-specific distribution of gap junctions defines cellular coupling to establish a vascular relay in the retina

    doi: 10.1002/cne.24699

    Figure Lengend Snippet: Validation and optimization of Connexin antibodies used in this study. (a–d) Cx37 labeling revealed just a few puncta (green) on blood vessels (blue) in pure PFA fixative (a) and entire outlines of contractile cells in carbodiimide-based fixatives (b and c). Ganglion cells were more visible in PFA fixative (asterisks); the lowest background was in carbodiimide. Pre-absorption of the antibody with a control peptide eliminated the labeling (d). (e–h) Cx40 (green) was associated with outlines of endothelial cells (blue) in all fixative with the less background in carbodiimide (f). Incubation of primary antibody eliminated the labeling (h) with the exception of labeled microglia (arrow). (i–k) Cx43s-positive puncta (green, Sigma antibody) were associated with vasculature in PFA fixative (i); in addition string-like structures along endothelial cell contacts (blue) became visible in carbodiimide fixative (j–k). (l) Omission of the primary antibody eliminated punctate and string-like structures; the microglia was labeled by the secondary antibody (arrow). (m–p) Cx43a produced identical pattern as Cx43s in carbodiimide containing fixatives (n, o). In pure PFA fixative, this antibody labeled well punctate and string-like structures; however the background was significantly higher (m). Incubation of the primary antibodies with control peptide eliminated the specific labeling (p). (q–s) Morphology of DsRed expressing pericytes was best preserved in PFA-containing fixatives (q and s); the DsRed labeling in pericyte processes was lost in pure carbodiimide fixative (q). (t) Summary of fixation procedure. Red crosses indicate signal, black crosses – background; the number of crosses indicates intensity. Scale bar 10 μm. Cx43s – Cx43 antibody from Sigma, Cx43a – Cx43 antibody from Alomone, NB – Neurobiotin.

    Article Snippet: All connexin antibodies from Alomone came with control peptides.

    Techniques: Labeling, Incubation, Produced, Expressing

    Brainstem Cx36 expression decreases because of nicotine or carbenoxolone application. ( a ) Example of western immunoblotting (top) showing the expression of Cx36 in brainstems incubated for 4 h in Krebs, or treated with TBOA, nicotine + TBOA, nicotine, or carbenoxolone + TBOA. Histograms (bottom) quantifying the significantly decreased Cx36 levels after treatment with nicotine + TBOA (n, N =9) or carbenoxolone + TBOA (n, N =5; Kruskal–Wallis one-way ANOVA: P =0.015 among groups). ( b and c ) Example of western blot (top) showing Cx36 expression in membrane ( b ) or the cytoplasmic ( c ) fractions of samples treated as described above. Histograms illustrate how nicotine alone or co-applied induces a significant decrease of Cx36 expression in the membrane pool ( b , bottom; Kruskal–Wallis one-way ANOVA: P =0.069 among groups; Mann–Whitney test: * P =0.016 for sham versus nicotine + TBOA and ** P =0.008 for sham versus nicotine; n, N =5) and an increase in the cytoplasmic one ( c , bottom; Kruskal–Wallis one-way ANOVA: P =0.014 among groups; Tukey test: * P

    Journal: Cell Death & Disease

    Article Title: Nicotine protects rat hypoglossal motoneurons from excitotoxic death via downregulation of connexin 36

    doi: 10.1038/cddis.2017.232

    Figure Lengend Snippet: Brainstem Cx36 expression decreases because of nicotine or carbenoxolone application. ( a ) Example of western immunoblotting (top) showing the expression of Cx36 in brainstems incubated for 4 h in Krebs, or treated with TBOA, nicotine + TBOA, nicotine, or carbenoxolone + TBOA. Histograms (bottom) quantifying the significantly decreased Cx36 levels after treatment with nicotine + TBOA (n, N =9) or carbenoxolone + TBOA (n, N =5; Kruskal–Wallis one-way ANOVA: P =0.015 among groups). ( b and c ) Example of western blot (top) showing Cx36 expression in membrane ( b ) or the cytoplasmic ( c ) fractions of samples treated as described above. Histograms illustrate how nicotine alone or co-applied induces a significant decrease of Cx36 expression in the membrane pool ( b , bottom; Kruskal–Wallis one-way ANOVA: P =0.069 among groups; Mann–Whitney test: * P =0.016 for sham versus nicotine + TBOA and ** P =0.008 for sham versus nicotine; n, N =5) and an increase in the cytoplasmic one ( c , bottom; Kruskal–Wallis one-way ANOVA: P =0.014 among groups; Tukey test: * P

    Article Snippet: Samples were then immunoblotted with rabbit anti-Cx36 (1:200, Alomone), rabbit anti-Panx1 (1:400, Alomone), mouse anti-Hsp70 (1:5000, Abcam, Cambridge, UK), mouse anti-β -actin (1:2000, Sigma-Aldrich), mouse anti-β -tubulin (1:2000, Sigma-Aldrich), or mouse anti-synaptophysin (1:10 000, Millipore) antibodies.

    Techniques: Expressing, Western Blot, Incubation, MANN-WHITNEY

    Cx36 mRNA levels and Hsp70 expression. ( a ) Plot illustrating unchanged values of Cx36 mRNA levels among different treatments. Fractional Cx36 expression (AU): 1±0.0, sham; 0.9±0.2, TBOA; 1±0.3, nicotine + TBOA; 1.3±0.2, nicotine (n, N =6). ( b ) Example of Hsp70 immunoblotting. ( c ) Histogram quantifying the significant increase of Hsp70 expression after nicotine + TBOA treatment (Kruskal–Wallis one-way ANOVA: P =0.027 among groups; Holm–Sidak method: * P

    Journal: Cell Death & Disease

    Article Title: Nicotine protects rat hypoglossal motoneurons from excitotoxic death via downregulation of connexin 36

    doi: 10.1038/cddis.2017.232

    Figure Lengend Snippet: Cx36 mRNA levels and Hsp70 expression. ( a ) Plot illustrating unchanged values of Cx36 mRNA levels among different treatments. Fractional Cx36 expression (AU): 1±0.0, sham; 0.9±0.2, TBOA; 1±0.3, nicotine + TBOA; 1.3±0.2, nicotine (n, N =6). ( b ) Example of Hsp70 immunoblotting. ( c ) Histogram quantifying the significant increase of Hsp70 expression after nicotine + TBOA treatment (Kruskal–Wallis one-way ANOVA: P =0.027 among groups; Holm–Sidak method: * P

    Article Snippet: Samples were then immunoblotted with rabbit anti-Cx36 (1:200, Alomone), rabbit anti-Panx1 (1:400, Alomone), mouse anti-Hsp70 (1:5000, Abcam, Cambridge, UK), mouse anti-β -actin (1:2000, Sigma-Aldrich), mouse anti-β -tubulin (1:2000, Sigma-Aldrich), or mouse anti-synaptophysin (1:10 000, Millipore) antibodies.

    Techniques: Expressing

    Idealized diagram to account for the toxic effects by TBOA and their antagonism by nicotine on HMs. ( a ) In the presence of TBOA, glutamate (filled red circles) released from presynaptic terminals acts on glutamatergic receptors to depolarize HMs. TBOA-mediated inhibition of glutamate uptake by astrocytes leads to intensification of glutamate effects with build-up of intracellular free Ca 2+ , which acts on mitochondria and favors production of ROS, release of AIF and DNA damage. This process is greatly amplified by the bidirectional (red arrows) operation of gap junctions (Cx36; filled blue cylinders) that spread and recruit HMs into a hyperexcitable state with deleterious consequences on cell survival. ( b ) In the presence of nicotine, the neurotoxic effect of TBOA is largely attenuated. Nicotine exerts multiple effects via nAChRs located on presynaptical terminals (with consequent decrease in glutamate release, and increase in the inhibitory response), HM membrane and even mitochondria. Through these effects, nicotine strongly decreases the motoneuron coupling via gap junctions through redistribution of Cx36 subunits, therefore disjoining motoneurons from their collective excitatory behavior. Our hypothesis is that this process is accompanied by inhibition of Cx36 activity, reduction in production of ROS and facilitation of the effect of Hsp70 to sequester AIF and prevent its nuclear migration. For references, see text

    Journal: Cell Death & Disease

    Article Title: Nicotine protects rat hypoglossal motoneurons from excitotoxic death via downregulation of connexin 36

    doi: 10.1038/cddis.2017.232

    Figure Lengend Snippet: Idealized diagram to account for the toxic effects by TBOA and their antagonism by nicotine on HMs. ( a ) In the presence of TBOA, glutamate (filled red circles) released from presynaptic terminals acts on glutamatergic receptors to depolarize HMs. TBOA-mediated inhibition of glutamate uptake by astrocytes leads to intensification of glutamate effects with build-up of intracellular free Ca 2+ , which acts on mitochondria and favors production of ROS, release of AIF and DNA damage. This process is greatly amplified by the bidirectional (red arrows) operation of gap junctions (Cx36; filled blue cylinders) that spread and recruit HMs into a hyperexcitable state with deleterious consequences on cell survival. ( b ) In the presence of nicotine, the neurotoxic effect of TBOA is largely attenuated. Nicotine exerts multiple effects via nAChRs located on presynaptical terminals (with consequent decrease in glutamate release, and increase in the inhibitory response), HM membrane and even mitochondria. Through these effects, nicotine strongly decreases the motoneuron coupling via gap junctions through redistribution of Cx36 subunits, therefore disjoining motoneurons from their collective excitatory behavior. Our hypothesis is that this process is accompanied by inhibition of Cx36 activity, reduction in production of ROS and facilitation of the effect of Hsp70 to sequester AIF and prevent its nuclear migration. For references, see text

    Article Snippet: Samples were then immunoblotted with rabbit anti-Cx36 (1:200, Alomone), rabbit anti-Panx1 (1:400, Alomone), mouse anti-Hsp70 (1:5000, Abcam, Cambridge, UK), mouse anti-β -actin (1:2000, Sigma-Aldrich), mouse anti-β -tubulin (1:2000, Sigma-Aldrich), or mouse anti-synaptophysin (1:10 000, Millipore) antibodies.

    Techniques: Inhibition, Amplification, Activity Assay, Migration

    Nicotine and carbenoxolone induce significant reduction in Cx36 expression. ( a ) Example of HMs after 4-h incubation in Krebs (sham), TBOA, nicotine + TBOA, nicotine, or carbenoxolone + TBOA. HMs were labeled with SMI 32 (green pseudocolor; left column), as motoneuronal marker, or Cx36 (red pseudocolor; middle column). The right column shows merged images where DAPI (blue pseudocolor) is used to visualize nuclei. ( b ) Histograms quantify the Cx36 signal decrease after 4h of nicotine + TBOA (Kruskal–Wallis one-way ANOVA: P =0.005 among groups; Dunn’s method: * P

    Journal: Cell Death & Disease

    Article Title: Nicotine protects rat hypoglossal motoneurons from excitotoxic death via downregulation of connexin 36

    doi: 10.1038/cddis.2017.232

    Figure Lengend Snippet: Nicotine and carbenoxolone induce significant reduction in Cx36 expression. ( a ) Example of HMs after 4-h incubation in Krebs (sham), TBOA, nicotine + TBOA, nicotine, or carbenoxolone + TBOA. HMs were labeled with SMI 32 (green pseudocolor; left column), as motoneuronal marker, or Cx36 (red pseudocolor; middle column). The right column shows merged images where DAPI (blue pseudocolor) is used to visualize nuclei. ( b ) Histograms quantify the Cx36 signal decrease after 4h of nicotine + TBOA (Kruskal–Wallis one-way ANOVA: P =0.005 among groups; Dunn’s method: * P

    Article Snippet: Samples were then immunoblotted with rabbit anti-Cx36 (1:200, Alomone), rabbit anti-Panx1 (1:400, Alomone), mouse anti-Hsp70 (1:5000, Abcam, Cambridge, UK), mouse anti-β -actin (1:2000, Sigma-Aldrich), mouse anti-β -tubulin (1:2000, Sigma-Aldrich), or mouse anti-synaptophysin (1:10 000, Millipore) antibodies.

    Techniques: Expressing, Incubation, Labeling, Marker

    Expression profile of CX45, CX36, and CX26 transcripts in the human fetal cortex, from 17 to 23 GW. ( A ) Log10 FPKM values derived by the HISAT-StringTie pathway of data analysis. ( B ) Log10 normalized counts derived by the STAR-HTSeq-DESeq2 data analysis pathway. ( C ) Expression profile of 2 predominant transcripts of Cx45 (ENST00000592524.5 [CX45_524.5] and ENST00000426548.5 [Cx45_548.5]) in comparison with Cx36 and Cx26 with HISAT-StringTie pathway of analysis from 17 to 23 GW in the human fetal cortex. In panels A – C , n = 4, 17 GW; n = 2, 18 GW; n = 5, 19 GW; and n = 1, 23 GW. ( D ) Log10 normalized counts of the Cx and pannexin isoforms expressed in the human fetal cortex, age range of 17–23 GW (STAR-HTSeq-DESeq2 data analysis pathway) ( n = 7). ( E ) Differential expression analysis of human fetal samples at 17 and 19 GW, by the STAR-HTSeq-DESeq2 method ( n = 3, 17 GW; n = 2, 19 GW). ( F ) Sex determination based on XIST expression and ratio of X-to-Y coverage in each human subject used in the study ( n = 7). ( G ) Differential expression analysis of male ( n = 4) and female ( n = 3) human fetal samples from 17 to 19 GW, by the STAR-HTSeq-DESeq2 method. ( H ) Differential expression analysis of cortex ( n = 3) and full brain ( n = 3) human fetal samples at 17 GW by the STAR-HTSeq-DESeq2 method. * P adj

    Journal: Cerebral Cortex (New York, NY)

    Article Title: Cadmium versus Lanthanum Effects on Spontaneous Electrical Activity and Expression of Connexin Isoforms Cx26, Cx36, and Cx45 in the Human Fetal Cortex

    doi: 10.1093/cercor/bhz163

    Figure Lengend Snippet: Expression profile of CX45, CX36, and CX26 transcripts in the human fetal cortex, from 17 to 23 GW. ( A ) Log10 FPKM values derived by the HISAT-StringTie pathway of data analysis. ( B ) Log10 normalized counts derived by the STAR-HTSeq-DESeq2 data analysis pathway. ( C ) Expression profile of 2 predominant transcripts of Cx45 (ENST00000592524.5 [CX45_524.5] and ENST00000426548.5 [Cx45_548.5]) in comparison with Cx36 and Cx26 with HISAT-StringTie pathway of analysis from 17 to 23 GW in the human fetal cortex. In panels A – C , n = 4, 17 GW; n = 2, 18 GW; n = 5, 19 GW; and n = 1, 23 GW. ( D ) Log10 normalized counts of the Cx and pannexin isoforms expressed in the human fetal cortex, age range of 17–23 GW (STAR-HTSeq-DESeq2 data analysis pathway) ( n = 7). ( E ) Differential expression analysis of human fetal samples at 17 and 19 GW, by the STAR-HTSeq-DESeq2 method ( n = 3, 17 GW; n = 2, 19 GW). ( F ) Sex determination based on XIST expression and ratio of X-to-Y coverage in each human subject used in the study ( n = 7). ( G ) Differential expression analysis of male ( n = 4) and female ( n = 3) human fetal samples from 17 to 19 GW, by the STAR-HTSeq-DESeq2 method. ( H ) Differential expression analysis of cortex ( n = 3) and full brain ( n = 3) human fetal samples at 17 GW by the STAR-HTSeq-DESeq2 method. * P adj

    Article Snippet: Primary antibodies were purchased from Alomone Labs, Thermo Fisher Scientific, and Millipore (company, antigen [catalog number]: Alomone Labs, Cx26 [ACC-212], Cx36 [ACC-209], and Cx45 [ACC-207]; Thermo Fisher, Cx26 [512800], Cx36 [516200], Nurr1 [MA1-195], beta-3-tubulin [MA1-19582], and NeuN [MA5-33103]; Millipore, NeuN [MAB377]).

    Techniques: Expressing, Derivative Assay

    Cells expressing Cx26 in 3 zones of cortical mantle. ( A ) Human fetal cortex (20 GW, female). Hoechst stain. Composite (mosaic) of multiple images. ( B ) CP, ( C ) SP, and ( D ) SVZ. Green = neuronal marker, Neun; red = Cx isoform, Cx26; blue = nuclear stain by Hoechst. Scales in panels B – D , 20 μm.

    Journal: Cerebral Cortex (New York, NY)

    Article Title: Cadmium versus Lanthanum Effects on Spontaneous Electrical Activity and Expression of Connexin Isoforms Cx26, Cx36, and Cx45 in the Human Fetal Cortex

    doi: 10.1093/cercor/bhz163

    Figure Lengend Snippet: Cells expressing Cx26 in 3 zones of cortical mantle. ( A ) Human fetal cortex (20 GW, female). Hoechst stain. Composite (mosaic) of multiple images. ( B ) CP, ( C ) SP, and ( D ) SVZ. Green = neuronal marker, Neun; red = Cx isoform, Cx26; blue = nuclear stain by Hoechst. Scales in panels B – D , 20 μm.

    Article Snippet: Primary antibodies were purchased from Alomone Labs, Thermo Fisher Scientific, and Millipore (company, antigen [catalog number]: Alomone Labs, Cx26 [ACC-212], Cx36 [ACC-209], and Cx45 [ACC-207]; Thermo Fisher, Cx26 [512800], Cx36 [516200], Nurr1 [MA1-195], beta-3-tubulin [MA1-19582], and NeuN [MA5-33103]; Millipore, NeuN [MAB377]).

    Techniques: Expressing, Staining, Marker