ncx3  (Alomone Labs)


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    Alomone Labs ncx3
    Impact of bepridil on human astrocytes (HA) and the Na + /Ca 2+ exchanger (NCX) isoforms in each cell line. (a) Viability of U87, U251, and SF188 cells and HA after exposure to bepridil (25 μM, 48 hr); n = 5 independent tests in each group. (b) The bands of NCX1, NCX2, and <t>NCX3</t> isoforms detected by Western blot analysis in the sample of HA and glioblastoma cell lines U87, U118, A172, U251, and SF188. (c) Representative recording of the NCX currents in HA and glioblastoma cell lines. (d) Viability of U87, U251, and SF188 cells and HA after exposure to KB‐R7943 (25 μM, 48 hr). n = 5 independent tests in each group
    Ncx3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ncx3/product/Alomone Labs
    Average 94 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    ncx3 - by Bioz Stars, 2022-08
    94/100 stars

    Images

    1) Product Images from "Blockade of the forward Na+/Ca2+ exchanger suppresses the growth of glioblastoma cells through Ca2+‐mediated cell death, et al. Blockade of the forward Na+/Ca2+ exchanger suppresses the growth of glioblastoma cells through Ca2+‐mediated cell death"

    Article Title: Blockade of the forward Na+/Ca2+ exchanger suppresses the growth of glioblastoma cells through Ca2+‐mediated cell death, et al. Blockade of the forward Na+/Ca2+ exchanger suppresses the growth of glioblastoma cells through Ca2+‐mediated cell death

    Journal: British Journal of Pharmacology

    doi: 10.1111/bph.14692

    Impact of bepridil on human astrocytes (HA) and the Na + /Ca 2+ exchanger (NCX) isoforms in each cell line. (a) Viability of U87, U251, and SF188 cells and HA after exposure to bepridil (25 μM, 48 hr); n = 5 independent tests in each group. (b) The bands of NCX1, NCX2, and NCX3 isoforms detected by Western blot analysis in the sample of HA and glioblastoma cell lines U87, U118, A172, U251, and SF188. (c) Representative recording of the NCX currents in HA and glioblastoma cell lines. (d) Viability of U87, U251, and SF188 cells and HA after exposure to KB‐R7943 (25 μM, 48 hr). n = 5 independent tests in each group
    Figure Legend Snippet: Impact of bepridil on human astrocytes (HA) and the Na + /Ca 2+ exchanger (NCX) isoforms in each cell line. (a) Viability of U87, U251, and SF188 cells and HA after exposure to bepridil (25 μM, 48 hr); n = 5 independent tests in each group. (b) The bands of NCX1, NCX2, and NCX3 isoforms detected by Western blot analysis in the sample of HA and glioblastoma cell lines U87, U118, A172, U251, and SF188. (c) Representative recording of the NCX currents in HA and glioblastoma cell lines. (d) Viability of U87, U251, and SF188 cells and HA after exposure to KB‐R7943 (25 μM, 48 hr). n = 5 independent tests in each group

    Techniques Used: Western Blot

    2) Product Images from "Increased Piezo1 channel activity in interstitial Cajal-like cells induces bladder hyperactivity by functionally interacting with NCX1 in rats with cyclophosphamide-induced cystitis"

    Article Title: Increased Piezo1 channel activity in interstitial Cajal-like cells induces bladder hyperactivity by functionally interacting with NCX1 in rats with cyclophosphamide-induced cystitis

    Journal: Experimental & Molecular Medicine

    doi: 10.1038/s12276-018-0088-z

    Na + /Ca 2+ exchanger (NCX) expression levels were significantly increased and the reverse mode of NCX was relatively activated in bladder ICC-LCs from CYP-8d rats. a – c The mRNA and protein expression levels of NCX1 and NCX3 were significantly increased in CYP-48h and CYP-8d rat bladders compared with those in naive and CYP-4h rat bladders, and increases in CYP-8d rat bladders were more significant; the mRNA and protein expression levels of NCX2 were only increased in CYP-8d rat bladders. d Changes in NCX1 expression (red) in bladder c-kit (green)-positive ICC-LCs (white arrows) were detected using IF staining. Bladder ICC-LCs from CYP-8d rats possessed higher NCX1 protein expression than those from naive rats. e , f NCX current (I NCX ) in isolated bladder ICC-LCs from naive and CYP-8d rats was evoked by a step potential ranging from −100 to +60 mV in increments of 10 mV for 200 ms, with a holding potential of −40 mV. The forward and reverse modes of NCX were elicited over a voltage range of −100 to −50 mV and −30 to +60 mV, respectively, and are exhibited as the bottom and upper part of the characteristic I NCX traces. When normalized to cell capacitance, the absolute value of the I NCX density in bladder ICC-LCs from CYP-8d rats significantly exceeded that in bladder ICC-LCs from naive rats over a voltage range of −100 to −80 mV (forward mode) and +10 to +60 mV (reverse mode). Notably, the increase in the reverse mode of NCX was more significant (the data represent the mean ± S.D., n = 8; * P
    Figure Legend Snippet: Na + /Ca 2+ exchanger (NCX) expression levels were significantly increased and the reverse mode of NCX was relatively activated in bladder ICC-LCs from CYP-8d rats. a – c The mRNA and protein expression levels of NCX1 and NCX3 were significantly increased in CYP-48h and CYP-8d rat bladders compared with those in naive and CYP-4h rat bladders, and increases in CYP-8d rat bladders were more significant; the mRNA and protein expression levels of NCX2 were only increased in CYP-8d rat bladders. d Changes in NCX1 expression (red) in bladder c-kit (green)-positive ICC-LCs (white arrows) were detected using IF staining. Bladder ICC-LCs from CYP-8d rats possessed higher NCX1 protein expression than those from naive rats. e , f NCX current (I NCX ) in isolated bladder ICC-LCs from naive and CYP-8d rats was evoked by a step potential ranging from −100 to +60 mV in increments of 10 mV for 200 ms, with a holding potential of −40 mV. The forward and reverse modes of NCX were elicited over a voltage range of −100 to −50 mV and −30 to +60 mV, respectively, and are exhibited as the bottom and upper part of the characteristic I NCX traces. When normalized to cell capacitance, the absolute value of the I NCX density in bladder ICC-LCs from CYP-8d rats significantly exceeded that in bladder ICC-LCs from naive rats over a voltage range of −100 to −80 mV (forward mode) and +10 to +60 mV (reverse mode). Notably, the increase in the reverse mode of NCX was more significant (the data represent the mean ± S.D., n = 8; * P

    Techniques Used: Expressing, Immunocytochemistry, Staining, Isolation, Mass Spectrometry

    3) Product Images from "Increased Piezo1 channel activity in interstitial Cajal-like cells induces bladder hyperactivity by functionally interacting with NCX1 in rats with cyclophosphamide-induced cystitis"

    Article Title: Increased Piezo1 channel activity in interstitial Cajal-like cells induces bladder hyperactivity by functionally interacting with NCX1 in rats with cyclophosphamide-induced cystitis

    Journal: Experimental & Molecular Medicine

    doi: 10.1038/s12276-018-0088-z

    Na + /Ca 2+ exchanger (NCX) expression levels were significantly increased and the reverse mode of NCX was relatively activated in bladder ICC-LCs from CYP-8d rats. a – c The mRNA and protein expression levels of NCX1 and NCX3 were significantly increased in CYP-48h and CYP-8d rat bladders compared with those in naive and CYP-4h rat bladders, and increases in CYP-8d rat bladders were more significant; the mRNA and protein expression levels of NCX2 were only increased in CYP-8d rat bladders. d Changes in NCX1 expression (red) in bladder c-kit (green)-positive ICC-LCs (white arrows) were detected using IF staining. Bladder ICC-LCs from CYP-8d rats possessed higher NCX1 protein expression than those from naive rats. e , f NCX current (I NCX ) in isolated bladder ICC-LCs from naive and CYP-8d rats was evoked by a step potential ranging from −100 to +60 mV in increments of 10 mV for 200 ms, with a holding potential of −40 mV. The forward and reverse modes of NCX were elicited over a voltage range of −100 to −50 mV and −30 to +60 mV, respectively, and are exhibited as the bottom and upper part of the characteristic I NCX traces. When normalized to cell capacitance, the absolute value of the I NCX density in bladder ICC-LCs from CYP-8d rats significantly exceeded that in bladder ICC-LCs from naive rats over a voltage range of −100 to −80 mV (forward mode) and +10 to +60 mV (reverse mode). Notably, the increase in the reverse mode of NCX was more significant (the data represent the mean ± S.D., n = 8; * P
    Figure Legend Snippet: Na + /Ca 2+ exchanger (NCX) expression levels were significantly increased and the reverse mode of NCX was relatively activated in bladder ICC-LCs from CYP-8d rats. a – c The mRNA and protein expression levels of NCX1 and NCX3 were significantly increased in CYP-48h and CYP-8d rat bladders compared with those in naive and CYP-4h rat bladders, and increases in CYP-8d rat bladders were more significant; the mRNA and protein expression levels of NCX2 were only increased in CYP-8d rat bladders. d Changes in NCX1 expression (red) in bladder c-kit (green)-positive ICC-LCs (white arrows) were detected using IF staining. Bladder ICC-LCs from CYP-8d rats possessed higher NCX1 protein expression than those from naive rats. e , f NCX current (I NCX ) in isolated bladder ICC-LCs from naive and CYP-8d rats was evoked by a step potential ranging from −100 to +60 mV in increments of 10 mV for 200 ms, with a holding potential of −40 mV. The forward and reverse modes of NCX were elicited over a voltage range of −100 to −50 mV and −30 to +60 mV, respectively, and are exhibited as the bottom and upper part of the characteristic I NCX traces. When normalized to cell capacitance, the absolute value of the I NCX density in bladder ICC-LCs from CYP-8d rats significantly exceeded that in bladder ICC-LCs from naive rats over a voltage range of −100 to −80 mV (forward mode) and +10 to +60 mV (reverse mode). Notably, the increase in the reverse mode of NCX was more significant (the data represent the mean ± S.D., n = 8; * P

    Techniques Used: Expressing, Immunocytochemistry, Staining, Isolation, Mass Spectrometry

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    Alomone Labs ncx3
    Impact of bepridil on human astrocytes (HA) and the Na + /Ca 2+ exchanger (NCX) isoforms in each cell line. (a) Viability of U87, U251, and SF188 cells and HA after exposure to bepridil (25 μM, 48 hr); n = 5 independent tests in each group. (b) The bands of NCX1, NCX2, and <t>NCX3</t> isoforms detected by Western blot analysis in the sample of HA and glioblastoma cell lines U87, U118, A172, U251, and SF188. (c) Representative recording of the NCX currents in HA and glioblastoma cell lines. (d) Viability of U87, U251, and SF188 cells and HA after exposure to KB‐R7943 (25 μM, 48 hr). n = 5 independent tests in each group
    Ncx3, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ncx3/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ncx3 - by Bioz Stars, 2022-08
    94/100 stars
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    Impact of bepridil on human astrocytes (HA) and the Na + /Ca 2+ exchanger (NCX) isoforms in each cell line. (a) Viability of U87, U251, and SF188 cells and HA after exposure to bepridil (25 μM, 48 hr); n = 5 independent tests in each group. (b) The bands of NCX1, NCX2, and NCX3 isoforms detected by Western blot analysis in the sample of HA and glioblastoma cell lines U87, U118, A172, U251, and SF188. (c) Representative recording of the NCX currents in HA and glioblastoma cell lines. (d) Viability of U87, U251, and SF188 cells and HA after exposure to KB‐R7943 (25 μM, 48 hr). n = 5 independent tests in each group

    Journal: British Journal of Pharmacology

    Article Title: Blockade of the forward Na+/Ca2+ exchanger suppresses the growth of glioblastoma cells through Ca2+‐mediated cell death, et al. Blockade of the forward Na+/Ca2+ exchanger suppresses the growth of glioblastoma cells through Ca2+‐mediated cell death

    doi: 10.1111/bph.14692

    Figure Lengend Snippet: Impact of bepridil on human astrocytes (HA) and the Na + /Ca 2+ exchanger (NCX) isoforms in each cell line. (a) Viability of U87, U251, and SF188 cells and HA after exposure to bepridil (25 μM, 48 hr); n = 5 independent tests in each group. (b) The bands of NCX1, NCX2, and NCX3 isoforms detected by Western blot analysis in the sample of HA and glioblastoma cell lines U87, U118, A172, U251, and SF188. (c) Representative recording of the NCX currents in HA and glioblastoma cell lines. (d) Viability of U87, U251, and SF188 cells and HA after exposure to KB‐R7943 (25 μM, 48 hr). n = 5 independent tests in each group

    Article Snippet: Anti‐NCX1 antibody (Cat# ab177952, RRID:AB_2801276) was purchased from Abcam (Cambridge, MA, USA), and anti‐NCX2 (Cat# ANX‐012, RRID:AB_2341022) and NCX3 (Cat# ANX‐013, RRID:AB_2341023) antibodies were purchased from Alomone Labs (Jerusalem, Israel). http://www.jkchemical.com/CH/products/A01433521.html J & K Scientific (Sunnyvale, CA, USA).

    Techniques: Western Blot

    Na + /Ca 2+ exchanger (NCX) expression levels were significantly increased and the reverse mode of NCX was relatively activated in bladder ICC-LCs from CYP-8d rats. a – c The mRNA and protein expression levels of NCX1 and NCX3 were significantly increased in CYP-48h and CYP-8d rat bladders compared with those in naive and CYP-4h rat bladders, and increases in CYP-8d rat bladders were more significant; the mRNA and protein expression levels of NCX2 were only increased in CYP-8d rat bladders. d Changes in NCX1 expression (red) in bladder c-kit (green)-positive ICC-LCs (white arrows) were detected using IF staining. Bladder ICC-LCs from CYP-8d rats possessed higher NCX1 protein expression than those from naive rats. e , f NCX current (I NCX ) in isolated bladder ICC-LCs from naive and CYP-8d rats was evoked by a step potential ranging from −100 to +60 mV in increments of 10 mV for 200 ms, with a holding potential of −40 mV. The forward and reverse modes of NCX were elicited over a voltage range of −100 to −50 mV and −30 to +60 mV, respectively, and are exhibited as the bottom and upper part of the characteristic I NCX traces. When normalized to cell capacitance, the absolute value of the I NCX density in bladder ICC-LCs from CYP-8d rats significantly exceeded that in bladder ICC-LCs from naive rats over a voltage range of −100 to −80 mV (forward mode) and +10 to +60 mV (reverse mode). Notably, the increase in the reverse mode of NCX was more significant (the data represent the mean ± S.D., n = 8; * P

    Journal: Experimental & Molecular Medicine

    Article Title: Increased Piezo1 channel activity in interstitial Cajal-like cells induces bladder hyperactivity by functionally interacting with NCX1 in rats with cyclophosphamide-induced cystitis

    doi: 10.1038/s12276-018-0088-z

    Figure Lengend Snippet: Na + /Ca 2+ exchanger (NCX) expression levels were significantly increased and the reverse mode of NCX was relatively activated in bladder ICC-LCs from CYP-8d rats. a – c The mRNA and protein expression levels of NCX1 and NCX3 were significantly increased in CYP-48h and CYP-8d rat bladders compared with those in naive and CYP-4h rat bladders, and increases in CYP-8d rat bladders were more significant; the mRNA and protein expression levels of NCX2 were only increased in CYP-8d rat bladders. d Changes in NCX1 expression (red) in bladder c-kit (green)-positive ICC-LCs (white arrows) were detected using IF staining. Bladder ICC-LCs from CYP-8d rats possessed higher NCX1 protein expression than those from naive rats. e , f NCX current (I NCX ) in isolated bladder ICC-LCs from naive and CYP-8d rats was evoked by a step potential ranging from −100 to +60 mV in increments of 10 mV for 200 ms, with a holding potential of −40 mV. The forward and reverse modes of NCX were elicited over a voltage range of −100 to −50 mV and −30 to +60 mV, respectively, and are exhibited as the bottom and upper part of the characteristic I NCX traces. When normalized to cell capacitance, the absolute value of the I NCX density in bladder ICC-LCs from CYP-8d rats significantly exceeded that in bladder ICC-LCs from naive rats over a voltage range of −100 to −80 mV (forward mode) and +10 to +60 mV (reverse mode). Notably, the increase in the reverse mode of NCX was more significant (the data represent the mean ± S.D., n = 8; * P

    Article Snippet: After being blocked with 5% skim milk dissolved in Tris-buffered saline at room temperature for 2 h, the membranes were incubated overnight at 4 °C with various primary antibodies targeting the following proteins: IL-6 (Abcam, Cambridge, UK, ab9324, 1:800), TNF-α (Abcam, ab199013, 1:500), Piezo1 (Alomone labs, Jerusalem, Israel, APC-087, 1:500), NCX1 (Abcam, ab2869, 1:800), NCX2 (Alomone labs, ANX-012, 1:500), NCX3 (Alomone labs, ANX-013, 1:500), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Beyotime, AG019, 1:1000), and α-tubulin (Beyotime, AT819, 1:1000).

    Techniques: Expressing, Immunocytochemistry, Staining, Isolation, Mass Spectrometry

    Effect of NCX3 silencing or inhibition by KB-R7943 in WT and Tg2576 primary hippocampal neurons. (A) Representative superimposed traces of I NCX in the reverse and forward modes of operation recorded in WT and WT plus siNCX3 (black traces), Tg2576 and Tg2576 plus siNCX3 (grey traces) primary hippocampal neurons at 12 DIV. (B) Quantification of I NCX in the reverse mode of operation represented in A, expressed as percentage of variation in comparison to WT. Values are expressed as mean ± SEM of 3 independent experimental sessions. (C) Representative Western blotting experiments and relative quantifications showing the effect of NCX3 silencing (siNCX3) on NCX3, NCX1, and NCX2 protein expression in primary hippocampal neurons (D) Representative superimposed traces of I NCX in the reverse and forward modes of operation recorded from WT and WT plus 0.5 μM kB-R7943 (black traces), Tg2576 and Tg2576 plus 0.5 μM kB-R7943 (grey traces) primary hippocampal neurons at 12 DIV. (E) Quantification of I NCX in the reverse mode of operation represented in D, expressed as percentage of variation in comparison to WT. Values are expressed as mean ± SEM of 3 independent experimental sessions. The number of cells used for each experimental condition is noted on the bars. (F, G) Representative Western blot of NCX3 protein expression and densitometric quantification of NCX3 truncated band in WT and Tg2576 primary hippocampal neurons at 12 DIV, represented as percentage of WT. Values are expressed as mean ± SEM of 3 independent experimental sessions. Statistical comparisons between groups were performed by one-way ANOVA followed by Newman-Keuls’ test. (* p

    Journal: Frontiers in Pharmacology

    Article Title: The Na+/Ca2+ Exchanger 3 Is Functionally Coupled With the NaV1.6 Voltage-Gated Channel and Promotes an Endoplasmic Reticulum Ca2+ Refilling in a Transgenic Model of Alzheimer’s Disease

    doi: 10.3389/fphar.2021.775271

    Figure Lengend Snippet: Effect of NCX3 silencing or inhibition by KB-R7943 in WT and Tg2576 primary hippocampal neurons. (A) Representative superimposed traces of I NCX in the reverse and forward modes of operation recorded in WT and WT plus siNCX3 (black traces), Tg2576 and Tg2576 plus siNCX3 (grey traces) primary hippocampal neurons at 12 DIV. (B) Quantification of I NCX in the reverse mode of operation represented in A, expressed as percentage of variation in comparison to WT. Values are expressed as mean ± SEM of 3 independent experimental sessions. (C) Representative Western blotting experiments and relative quantifications showing the effect of NCX3 silencing (siNCX3) on NCX3, NCX1, and NCX2 protein expression in primary hippocampal neurons (D) Representative superimposed traces of I NCX in the reverse and forward modes of operation recorded from WT and WT plus 0.5 μM kB-R7943 (black traces), Tg2576 and Tg2576 plus 0.5 μM kB-R7943 (grey traces) primary hippocampal neurons at 12 DIV. (E) Quantification of I NCX in the reverse mode of operation represented in D, expressed as percentage of variation in comparison to WT. Values are expressed as mean ± SEM of 3 independent experimental sessions. The number of cells used for each experimental condition is noted on the bars. (F, G) Representative Western blot of NCX3 protein expression and densitometric quantification of NCX3 truncated band in WT and Tg2576 primary hippocampal neurons at 12 DIV, represented as percentage of WT. Values are expressed as mean ± SEM of 3 independent experimental sessions. Statistical comparisons between groups were performed by one-way ANOVA followed by Newman-Keuls’ test. (* p

    Article Snippet: The nitrocellulose membranes were incubated with the following antibodies: rabbit-polyclonal anti-NCX3, anti-NCX1, anti-NCX2 (1:1,000, Alomone Labs, Israel) and anti-β-actin peroxidase (1:10,000, Sigma-Aldrich, Milan, Italy).

    Techniques: Inhibition, Western Blot, Expressing

    Effect of NCX3 inhibition by KB-R7943 on ER Ca 2+ content in WT and Tg2576 primary hippocampal neurons. (A) Representative superimposed traces of [Ca 2+ ] i measured in WT (black trace, N = 36) and Tg2576 (grey trace, N = 34) primary hippocampal neurons at 12 DIV, representative images in panel (A) . (B) Quantification of basal values of [Ca 2+ ] i in WT ( N = 36) and Tg2576 primary hippocampal neurons at 12 DIV ( N = 34). (C) ER Ca 2+ content quantified as [Ca 2+ ] i increase induced by Thapsigargin (Tg; 1 μM) and ATP (100 μM) in 0 μM Ca 2+ , and expressed as percentage of the effect observed in WT (considered as 100%). (D) AUCs of [Ca 2+ ] i calculated for (A) . (E) Quantification of ER Ca 2+ content in WT and Tg2576 primary hippocampal neurons at 12 DIV treated with KB-R7943 at 0.5 μM. Values are represented as percentage of respective controls, expressed as mean ± SEM of 3 independent experimental sessions. Statistical comparisons between groups were performed by one-way ANOVA followed by Newman-Keuls’ test. (* p

    Journal: Frontiers in Pharmacology

    Article Title: The Na+/Ca2+ Exchanger 3 Is Functionally Coupled With the NaV1.6 Voltage-Gated Channel and Promotes an Endoplasmic Reticulum Ca2+ Refilling in a Transgenic Model of Alzheimer’s Disease

    doi: 10.3389/fphar.2021.775271

    Figure Lengend Snippet: Effect of NCX3 inhibition by KB-R7943 on ER Ca 2+ content in WT and Tg2576 primary hippocampal neurons. (A) Representative superimposed traces of [Ca 2+ ] i measured in WT (black trace, N = 36) and Tg2576 (grey trace, N = 34) primary hippocampal neurons at 12 DIV, representative images in panel (A) . (B) Quantification of basal values of [Ca 2+ ] i in WT ( N = 36) and Tg2576 primary hippocampal neurons at 12 DIV ( N = 34). (C) ER Ca 2+ content quantified as [Ca 2+ ] i increase induced by Thapsigargin (Tg; 1 μM) and ATP (100 μM) in 0 μM Ca 2+ , and expressed as percentage of the effect observed in WT (considered as 100%). (D) AUCs of [Ca 2+ ] i calculated for (A) . (E) Quantification of ER Ca 2+ content in WT and Tg2576 primary hippocampal neurons at 12 DIV treated with KB-R7943 at 0.5 μM. Values are represented as percentage of respective controls, expressed as mean ± SEM of 3 independent experimental sessions. Statistical comparisons between groups were performed by one-way ANOVA followed by Newman-Keuls’ test. (* p

    Article Snippet: The nitrocellulose membranes were incubated with the following antibodies: rabbit-polyclonal anti-NCX3, anti-NCX1, anti-NCX2 (1:1,000, Alomone Labs, Israel) and anti-β-actin peroxidase (1:10,000, Sigma-Aldrich, Milan, Italy).

    Techniques: Inhibition

    Distribution of Na V 1.6 channels and NCX3 in Tg2576 primary hippocampal neurons. (A) Confocal microscopic images displaying the distribution of Na V 1.6 (red) and NCX3 (green) immunoreactivities in hippocampal neurons isolated from WT and Tg2576 mouse embryos and cultured for 12 DIV (scale bars: in a–f: 20 µm). (B) Confocal microscopic images displaying the distribution of Na V 1.6 (red) and NCX3 (green) immunoreactivities in Tg2576 primary hippocampal neurons at 12 DIV. Arrows in a-c point to the intense co-localization of Na V 1.6 and NCX3 immunostaining along neurites (scale bars: 20 µm). (C) Densitometric analysis of Na V 1.6 (left) and NCX3 (right) fluorescence intensities in WT and Tg2576 neurons at 12 DIV. The data are expressed in arbitrary units (* p

    Journal: Frontiers in Pharmacology

    Article Title: The Na+/Ca2+ Exchanger 3 Is Functionally Coupled With the NaV1.6 Voltage-Gated Channel and Promotes an Endoplasmic Reticulum Ca2+ Refilling in a Transgenic Model of Alzheimer’s Disease

    doi: 10.3389/fphar.2021.775271

    Figure Lengend Snippet: Distribution of Na V 1.6 channels and NCX3 in Tg2576 primary hippocampal neurons. (A) Confocal microscopic images displaying the distribution of Na V 1.6 (red) and NCX3 (green) immunoreactivities in hippocampal neurons isolated from WT and Tg2576 mouse embryos and cultured for 12 DIV (scale bars: in a–f: 20 µm). (B) Confocal microscopic images displaying the distribution of Na V 1.6 (red) and NCX3 (green) immunoreactivities in Tg2576 primary hippocampal neurons at 12 DIV. Arrows in a-c point to the intense co-localization of Na V 1.6 and NCX3 immunostaining along neurites (scale bars: 20 µm). (C) Densitometric analysis of Na V 1.6 (left) and NCX3 (right) fluorescence intensities in WT and Tg2576 neurons at 12 DIV. The data are expressed in arbitrary units (* p

    Article Snippet: The nitrocellulose membranes were incubated with the following antibodies: rabbit-polyclonal anti-NCX3, anti-NCX1, anti-NCX2 (1:1,000, Alomone Labs, Israel) and anti-β-actin peroxidase (1:10,000, Sigma-Aldrich, Milan, Italy).

    Techniques: Isolation, Cell Culture, Immunostaining, Fluorescence

    Na + /Ca 2+ exchanger (NCX) expression levels were significantly increased and the reverse mode of NCX was relatively activated in bladder ICC-LCs from CYP-8d rats. a – c The mRNA and protein expression levels of NCX1 and NCX3 were significantly increased in CYP-48h and CYP-8d rat bladders compared with those in naive and CYP-4h rat bladders, and increases in CYP-8d rat bladders were more significant; the mRNA and protein expression levels of NCX2 were only increased in CYP-8d rat bladders. d Changes in NCX1 expression (red) in bladder c-kit (green)-positive ICC-LCs (white arrows) were detected using IF staining. Bladder ICC-LCs from CYP-8d rats possessed higher NCX1 protein expression than those from naive rats. e , f NCX current (I NCX ) in isolated bladder ICC-LCs from naive and CYP-8d rats was evoked by a step potential ranging from −100 to +60 mV in increments of 10 mV for 200 ms, with a holding potential of −40 mV. The forward and reverse modes of NCX were elicited over a voltage range of −100 to −50 mV and −30 to +60 mV, respectively, and are exhibited as the bottom and upper part of the characteristic I NCX traces. When normalized to cell capacitance, the absolute value of the I NCX density in bladder ICC-LCs from CYP-8d rats significantly exceeded that in bladder ICC-LCs from naive rats over a voltage range of −100 to −80 mV (forward mode) and +10 to +60 mV (reverse mode). Notably, the increase in the reverse mode of NCX was more significant (the data represent the mean ± S.D., n = 8; * P

    Journal: Experimental & Molecular Medicine

    Article Title: Increased Piezo1 channel activity in interstitial Cajal-like cells induces bladder hyperactivity by functionally interacting with NCX1 in rats with cyclophosphamide-induced cystitis

    doi: 10.1038/s12276-018-0088-z

    Figure Lengend Snippet: Na + /Ca 2+ exchanger (NCX) expression levels were significantly increased and the reverse mode of NCX was relatively activated in bladder ICC-LCs from CYP-8d rats. a – c The mRNA and protein expression levels of NCX1 and NCX3 were significantly increased in CYP-48h and CYP-8d rat bladders compared with those in naive and CYP-4h rat bladders, and increases in CYP-8d rat bladders were more significant; the mRNA and protein expression levels of NCX2 were only increased in CYP-8d rat bladders. d Changes in NCX1 expression (red) in bladder c-kit (green)-positive ICC-LCs (white arrows) were detected using IF staining. Bladder ICC-LCs from CYP-8d rats possessed higher NCX1 protein expression than those from naive rats. e , f NCX current (I NCX ) in isolated bladder ICC-LCs from naive and CYP-8d rats was evoked by a step potential ranging from −100 to +60 mV in increments of 10 mV for 200 ms, with a holding potential of −40 mV. The forward and reverse modes of NCX were elicited over a voltage range of −100 to −50 mV and −30 to +60 mV, respectively, and are exhibited as the bottom and upper part of the characteristic I NCX traces. When normalized to cell capacitance, the absolute value of the I NCX density in bladder ICC-LCs from CYP-8d rats significantly exceeded that in bladder ICC-LCs from naive rats over a voltage range of −100 to −80 mV (forward mode) and +10 to +60 mV (reverse mode). Notably, the increase in the reverse mode of NCX was more significant (the data represent the mean ± S.D., n = 8; * P

    Article Snippet: After being blocked with 5% skim milk dissolved in Tris-buffered saline at room temperature for 2 h, the membranes were incubated overnight at 4 °C with various primary antibodies targeting the following proteins: IL-6 (Abcam, Cambridge, UK, ab9324, 1:800), TNF-α (Abcam, ab199013, 1:500), Piezo1 (Alomone labs, Jerusalem, Israel, APC-087, 1:500), NCX1 (Abcam, ab2869, 1:800), NCX2 (Alomone labs, ANX-012, 1:500), NCX3 (Alomone labs, ANX-013, 1:500), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Beyotime, AG019, 1:1000), and α-tubulin (Beyotime, AT819, 1:1000).

    Techniques: Expressing, Immunocytochemistry, Staining, Isolation, Mass Spectrometry