stim1  (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
  • 93

    Structured Review

    Alomone Labs stim1
    Increased TRPC1 and <t>STIM1</t> in MetS are abolished by exercise. ( A ) Representative agarose gel image quantifying TRPC1 mRNA using RT–PCR. ( B ) TRPC1 mRNA normalized to β-actin. STIM1 ( C ) and Orai1 ( D ) mRNA using quantitative RT–PCR
    Stim1, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/stim1/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    stim1 - by Bioz Stars, 2022-07
    93/100 stars

    Images

    1) Product Images from "Exercise training decreases store-operated Ca2+entry associated with metabolic syndrome and coronary atherosclerosis"

    Article Title: Exercise training decreases store-operated Ca2+entry associated with metabolic syndrome and coronary atherosclerosis

    Journal: Cardiovascular Research

    doi: 10.1093/cvr/cvp308

    Increased TRPC1 and STIM1 in MetS are abolished by exercise. ( A ) Representative agarose gel image quantifying TRPC1 mRNA using RT–PCR. ( B ) TRPC1 mRNA normalized to β-actin. STIM1 ( C ) and Orai1 ( D ) mRNA using quantitative RT–PCR
    Figure Legend Snippet: Increased TRPC1 and STIM1 in MetS are abolished by exercise. ( A ) Representative agarose gel image quantifying TRPC1 mRNA using RT–PCR. ( B ) TRPC1 mRNA normalized to β-actin. STIM1 ( C ) and Orai1 ( D ) mRNA using quantitative RT–PCR

    Techniques Used: Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

    2) Product Images from "Inhibition of Polyamine Biosynthesis Reverses Ca2+ Channel Remodeling in Colon Cancer Cells"

    Article Title: Inhibition of Polyamine Biosynthesis Reverses Ca2+ Channel Remodeling in Colon Cancer Cells

    Journal: Cancers

    doi: 10.3390/cancers11010083

    Effects of DFMO on the expression of proteins involved in SOCE in colon cancer HT29 cells. HT29 cells were treated with vehicle (control) or DFMO 5 mM, and then cells were lysed and subjected to Western blotting with antibodies against TRPC1, STIM1, STIM2, ORAI1, ORAI2 and ORAI3, followed by reprobing with anti-β-actin antibody for protein loading control. Bar graphs represent specific protein expression normalized to the β-actin content. Data are from n = 3 experiments (* p
    Figure Legend Snippet: Effects of DFMO on the expression of proteins involved in SOCE in colon cancer HT29 cells. HT29 cells were treated with vehicle (control) or DFMO 5 mM, and then cells were lysed and subjected to Western blotting with antibodies against TRPC1, STIM1, STIM2, ORAI1, ORAI2 and ORAI3, followed by reprobing with anti-β-actin antibody for protein loading control. Bar graphs represent specific protein expression normalized to the β-actin content. Data are from n = 3 experiments (* p

    Techniques Used: Expressing, Western Blot

    Effects of DFMO on the expression of genes coding for SOCE molecular players in HT29 cells. mRNA expression levels of selected genes were determined using qRT-PCR of extracts from control and DFMO-treated HT29 cells. β-actin was used as a reference. Data results are mean ± SEM from DFMO-treated cells relative to untreated cells Data are from n = 7, 7, 6, 6, 6, and 5 experiments for TRPC1 , STIM1 , STIM2 , ORAI1 , ORAI2 , and ORAI3 , respectively * p
    Figure Legend Snippet: Effects of DFMO on the expression of genes coding for SOCE molecular players in HT29 cells. mRNA expression levels of selected genes were determined using qRT-PCR of extracts from control and DFMO-treated HT29 cells. β-actin was used as a reference. Data results are mean ± SEM from DFMO-treated cells relative to untreated cells Data are from n = 7, 7, 6, 6, 6, and 5 experiments for TRPC1 , STIM1 , STIM2 , ORAI1 , ORAI2 , and ORAI3 , respectively * p

    Techniques Used: Expressing, Quantitative RT-PCR

    3) Product Images from "Calcium Independent Effect of Orai1 and STIM1 in Non-Hodgkin B Cell Lymphoma Dissemination"

    Article Title: Calcium Independent Effect of Orai1 and STIM1 in Non-Hodgkin B Cell Lymphoma Dissemination

    Journal: Cancers

    doi: 10.3390/cancers10110402

    Stromal Derived Factor 1 (SDF-1) provokes an intracellular Ca 2+ response in the HLY-1 diffuse large B cell lymphoma (DLBCL) cell line involving intracellular Ca 2+ pool mobilization and Orai1/STIM1 extracellular Ca 2+ influx. Ca 2+ responses to SDF-1 (100 ng/mL) were measured using Fluo2-LR-AM Ca 2+ dye and recorded by videomicroscopy (Zeiss LSM 510) using ×25 objective. Black arrows indicate SDF-1 addition. Each trace represents the response of one cell and data are representative of at least three independent experiments. Typical response of unique cell (peak or peak follow by sustained plateau phase) are present as example ( Ab ). Data were processed using GraphPad prism. ( A ) Pharmacological characterization of SDF-1-induced Ca 2+ increase. Cells were recorded in extracellular saline solution (HBSS) containing 2 mM Ca 2+ (2 Ca, ( Aa )) or in Ca 2+ -free HBSS (0 Ca, ( Ac )). Cells were pre-incubated with BTP2 ( Ae ) or GSK7975A ( Af ) at 10 µM for 30 min and recorded in 2 mM Ca 2+ HBSS containing inhibitors. ( Ad , Ag ) Histograms represent areas under curves (AUC) calculated, under various recording conditions, between the application time of SDF-1 and t = 2050 s, and normalized compared to control (2 Ca or shNT). Data are expressed as mean ± SEM, * p
    Figure Legend Snippet: Stromal Derived Factor 1 (SDF-1) provokes an intracellular Ca 2+ response in the HLY-1 diffuse large B cell lymphoma (DLBCL) cell line involving intracellular Ca 2+ pool mobilization and Orai1/STIM1 extracellular Ca 2+ influx. Ca 2+ responses to SDF-1 (100 ng/mL) were measured using Fluo2-LR-AM Ca 2+ dye and recorded by videomicroscopy (Zeiss LSM 510) using ×25 objective. Black arrows indicate SDF-1 addition. Each trace represents the response of one cell and data are representative of at least three independent experiments. Typical response of unique cell (peak or peak follow by sustained plateau phase) are present as example ( Ab ). Data were processed using GraphPad prism. ( A ) Pharmacological characterization of SDF-1-induced Ca 2+ increase. Cells were recorded in extracellular saline solution (HBSS) containing 2 mM Ca 2+ (2 Ca, ( Aa )) or in Ca 2+ -free HBSS (0 Ca, ( Ac )). Cells were pre-incubated with BTP2 ( Ae ) or GSK7975A ( Af ) at 10 µM for 30 min and recorded in 2 mM Ca 2+ HBSS containing inhibitors. ( Ad , Ag ) Histograms represent areas under curves (AUC) calculated, under various recording conditions, between the application time of SDF-1 and t = 2050 s, and normalized compared to control (2 Ca or shNT). Data are expressed as mean ± SEM, * p

    Techniques Used: Derivative Assay, Incubation

    Orai1 and STIM1 regulate basal and SDF-1-induced DLBCL cell migration in a Ca 2+ independent manner in vitro. Cell migration was assessed in 96-transwell chemotaxis chambers assay. Histograms represent mean ± SEM from at least 3 independent experiments, * p
    Figure Legend Snippet: Orai1 and STIM1 regulate basal and SDF-1-induced DLBCL cell migration in a Ca 2+ independent manner in vitro. Cell migration was assessed in 96-transwell chemotaxis chambers assay. Histograms represent mean ± SEM from at least 3 independent experiments, * p

    Techniques Used: Migration, In Vitro, Chemotaxis Assay

    STIM1, but not Ca 2+ , regulate DLBCL dissemination in vivo. ( A ) Effect of STIM1 under-expression on HLY-1 cell dissemination. ( B ) Effect of intraperitoneal injection of BTP2 (12 µg/kg) or vehicle, three times per week on HLY-1 cell dissemination. Images were captured with a Nikon Eclipse Ci microscope equipped with a Plan Fluor 10× 0.3 NA objective. Scale bars = 150 μm. Histograms represent the quantification of positive surface for HLA-ABC staining. All tissues were delimited and to evaluate the percentage of positive surface for HLA-ABC staining on tissue, thresholding on positive and negative staining was done using Mercator software. Data are represented as mean ± SEM (n = 10), * p
    Figure Legend Snippet: STIM1, but not Ca 2+ , regulate DLBCL dissemination in vivo. ( A ) Effect of STIM1 under-expression on HLY-1 cell dissemination. ( B ) Effect of intraperitoneal injection of BTP2 (12 µg/kg) or vehicle, three times per week on HLY-1 cell dissemination. Images were captured with a Nikon Eclipse Ci microscope equipped with a Plan Fluor 10× 0.3 NA objective. Scale bars = 150 μm. Histograms represent the quantification of positive surface for HLA-ABC staining. All tissues were delimited and to evaluate the percentage of positive surface for HLA-ABC staining on tissue, thresholding on positive and negative staining was done using Mercator software. Data are represented as mean ± SEM (n = 10), * p

    Techniques Used: In Vivo, Expressing, Injection, Microscopy, Staining, Negative Staining, Software

    Orai1 and STIM1 control DLBCL cell migration through RhoA activation, ROCK, and MLC phosphorylation. ( A ) SDF-1-induced SU-DHL-4 and HLY-1 cell migration is ROCK activation dependent. Cells incubated or not with SDF-1 (100 ng/mL) were pre-treated during 20 min in the presence or not of various inhibitors (FAK inhibitor 1 µM, PD98059 a MEK inhibitor 10 µM, Wortmannin a PI3K inhibitor 10 nM, AKT inhibitor 250 nM, Y27632 a ROCK inhibitor 1 µM). Data are represented as mean ± SEM of 3 independent experiments, * p
    Figure Legend Snippet: Orai1 and STIM1 control DLBCL cell migration through RhoA activation, ROCK, and MLC phosphorylation. ( A ) SDF-1-induced SU-DHL-4 and HLY-1 cell migration is ROCK activation dependent. Cells incubated or not with SDF-1 (100 ng/mL) were pre-treated during 20 min in the presence or not of various inhibitors (FAK inhibitor 1 µM, PD98059 a MEK inhibitor 10 µM, Wortmannin a PI3K inhibitor 10 nM, AKT inhibitor 250 nM, Y27632 a ROCK inhibitor 1 µM). Data are represented as mean ± SEM of 3 independent experiments, * p

    Techniques Used: Migration, Activation Assay, Incubation

    Orai1 and STIM1 expression is altered in extra-nodal DLBCL. ( A ) Representative immunofluorescent staining of STIM1 and Orai1 in normal lymph node and extra-nodal DLBCL under-expressing STIM1 or Orai1. Images were acquired using Leica DMI8 microscope equipped with a 40× oil immersion objective. TMA including samples from normal lymph node (n = 26), nodal (n = 43) and extra-nodal (n = 44) DLBCL was co-immunostained using Orai1 or STIM1 antibodies revealed by donkey anti-rabbit Alexa-488 (in green) and mouse anti-human CD20 and anti-human CD19 revealed by goat anti-mouse Alexa 532 (in red). Nuclei were stained with DAPI (in blue). Scale bar = 50 µm. ( B ) Quantification of STIM1 or Orai1 alteration in DLBCL samples. After acquisition of spot fluorescence on Icys laser scanning cytometer, a segmentation analysis base on phantoms was done to determine the percentage of CD20/CD19 positive cells expressing protein of interest for each spot. The stacked column charts illustrate the intensity grade for STIM1 or Orai1 expression in surgical specimens. * p
    Figure Legend Snippet: Orai1 and STIM1 expression is altered in extra-nodal DLBCL. ( A ) Representative immunofluorescent staining of STIM1 and Orai1 in normal lymph node and extra-nodal DLBCL under-expressing STIM1 or Orai1. Images were acquired using Leica DMI8 microscope equipped with a 40× oil immersion objective. TMA including samples from normal lymph node (n = 26), nodal (n = 43) and extra-nodal (n = 44) DLBCL was co-immunostained using Orai1 or STIM1 antibodies revealed by donkey anti-rabbit Alexa-488 (in green) and mouse anti-human CD20 and anti-human CD19 revealed by goat anti-mouse Alexa 532 (in red). Nuclei were stained with DAPI (in blue). Scale bar = 50 µm. ( B ) Quantification of STIM1 or Orai1 alteration in DLBCL samples. After acquisition of spot fluorescence on Icys laser scanning cytometer, a segmentation analysis base on phantoms was done to determine the percentage of CD20/CD19 positive cells expressing protein of interest for each spot. The stacked column charts illustrate the intensity grade for STIM1 or Orai1 expression in surgical specimens. * p

    Techniques Used: Expressing, Staining, Microscopy, Fluorescence, Cytometry

    4) Product Images from "Mast Cell CRF2 Suppresses Mast Cell Degranulation and Limits the Severity of Anaphylaxis and Stress-Induced Intestinal Permeability"

    Article Title: Mast Cell CRF2 Suppresses Mast Cell Degranulation and Limits the Severity of Anaphylaxis and Stress-Induced Intestinal Permeability

    Journal: The Journal of allergy and clinical immunology

    doi: 10.1016/j.jaci.2018.08.053

    CRF2−/− BMMCs exhibit heightened intracellular Ca 2+ store release and expression of SOCE channels. BMMCs derived from WT and CRF 2 −/− mice were loaded with Fluo 4, and intracellular Ca 2+ levels were measured following stimulation with IgE/DNP. A,B: Representative intracellular Ca 2+ traces for experiments conducted under Ca 2+ -replete (A) or Ca 2+ -free (1 mM EDTA; B) conditions. C: Mean peak change in fluorescence following IgE/DNP stimulus presented as ∆ peak fluorescence. D-I: Representative Western blots and densitometry analysis for STIM1 (D, G), TRPC1 (E, H), and Orai (F, I) in WT and CRF 2 −/− BMMCs. *Significance between groups was determined by an unpaired two-tailed t-test (C, G-I),*p
    Figure Legend Snippet: CRF2−/− BMMCs exhibit heightened intracellular Ca 2+ store release and expression of SOCE channels. BMMCs derived from WT and CRF 2 −/− mice were loaded with Fluo 4, and intracellular Ca 2+ levels were measured following stimulation with IgE/DNP. A,B: Representative intracellular Ca 2+ traces for experiments conducted under Ca 2+ -replete (A) or Ca 2+ -free (1 mM EDTA; B) conditions. C: Mean peak change in fluorescence following IgE/DNP stimulus presented as ∆ peak fluorescence. D-I: Representative Western blots and densitometry analysis for STIM1 (D, G), TRPC1 (E, H), and Orai (F, I) in WT and CRF 2 −/− BMMCs. *Significance between groups was determined by an unpaired two-tailed t-test (C, G-I),*p

    Techniques Used: Expressing, Derivative Assay, Mouse Assay, Fluorescence, Western Blot, Two Tailed Test

    5) Product Images from "Inhibition of Polyamine Biosynthesis Reverses Ca2+ Channel Remodeling in Colon Cancer Cells"

    Article Title: Inhibition of Polyamine Biosynthesis Reverses Ca2+ Channel Remodeling in Colon Cancer Cells

    Journal: Cancers

    doi: 10.3390/cancers11010083

    Effects of DFMO on the expression of proteins involved in SOCE in colon cancer HT29 cells. HT29 cells were treated with vehicle (control) or DFMO 5 mM, and then cells were lysed and subjected to Western blotting with antibodies against TRPC1, STIM1, STIM2, ORAI1, ORAI2 and ORAI3, followed by reprobing with anti-β-actin antibody for protein loading control. Bar graphs represent specific protein expression normalized to the β-actin content. Data are from n = 3 experiments (* p
    Figure Legend Snippet: Effects of DFMO on the expression of proteins involved in SOCE in colon cancer HT29 cells. HT29 cells were treated with vehicle (control) or DFMO 5 mM, and then cells were lysed and subjected to Western blotting with antibodies against TRPC1, STIM1, STIM2, ORAI1, ORAI2 and ORAI3, followed by reprobing with anti-β-actin antibody for protein loading control. Bar graphs represent specific protein expression normalized to the β-actin content. Data are from n = 3 experiments (* p

    Techniques Used: Expressing, Western Blot

    Effects of DFMO on the expression of genes coding for SOCE molecular players in HT29 cells. mRNA expression levels of selected genes were determined using qRT-PCR of extracts from control and DFMO-treated HT29 cells. β-actin was used as a reference. Data results are mean ± SEM from DFMO-treated cells relative to untreated cells Data are from n = 7, 7, 6, 6, 6, and 5 experiments for TRPC1 , STIM1 , STIM2 , ORAI1 , ORAI2 , and ORAI3 , respectively * p
    Figure Legend Snippet: Effects of DFMO on the expression of genes coding for SOCE molecular players in HT29 cells. mRNA expression levels of selected genes were determined using qRT-PCR of extracts from control and DFMO-treated HT29 cells. β-actin was used as a reference. Data results are mean ± SEM from DFMO-treated cells relative to untreated cells Data are from n = 7, 7, 6, 6, 6, and 5 experiments for TRPC1 , STIM1 , STIM2 , ORAI1 , ORAI2 , and ORAI3 , respectively * p

    Techniques Used: Expressing, Quantitative RT-PCR

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Alomone Labs rabbit anti stim1
    Subcellular localization and interaction of amnionless (AMN), <t>STIM1</t> and ORAIs in PTECs. PTECs were cotransfected with AMN-EYFP, STIM1-mCherry and CFP-ORAIs and fluorescence was examined before and after treatment with thapsigargin (TG, 1 µM) for 10 min using TIRF microscopy with objective ( × 100). a Before TG treatment (Pre-TG). b After store-depletion with TG (post-TG). The fluorescence distribution pattern and intensity (below each image) are shown for cells transfected with CFP-ORAI1, CFP-ORAI2, and CFP-ORAI3, respectively, plus AMN-EYFP and STMI1-mCherry. c Pictures show the localization of STIM1-mCherry, CFP-ORAI1 and AMN-EYFP at the apical and basal membrane of human PTECs after treatment with 1 µM TG for 10 min. d No significant clusters were formed before or after treatment with SERCA blocker TG (1 µM) in PTECs transfected with AMN-EYFP alone. Images were taken from Epi-Fluorescence (EPI-F) and TIRF microscopy. e Co-immunoprecipitation of STIM1, ORAI1 and AMN endogenously expressed in PTECs with/without TG treatment. f Colocalization of intracellular C terminus-truncated AMN (AMN1-183) with STIM1 in PTECs after Ca 2+ store depletion. Scale bars, 10 µm
    Rabbit Anti Stim1, 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/rabbit anti stim1/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti stim1 - by Bioz Stars, 2022-07
    94/100 stars
      Buy from Supplier

    Image Search Results


    Subcellular localization and interaction of amnionless (AMN), STIM1 and ORAIs in PTECs. PTECs were cotransfected with AMN-EYFP, STIM1-mCherry and CFP-ORAIs and fluorescence was examined before and after treatment with thapsigargin (TG, 1 µM) for 10 min using TIRF microscopy with objective ( × 100). a Before TG treatment (Pre-TG). b After store-depletion with TG (post-TG). The fluorescence distribution pattern and intensity (below each image) are shown for cells transfected with CFP-ORAI1, CFP-ORAI2, and CFP-ORAI3, respectively, plus AMN-EYFP and STMI1-mCherry. c Pictures show the localization of STIM1-mCherry, CFP-ORAI1 and AMN-EYFP at the apical and basal membrane of human PTECs after treatment with 1 µM TG for 10 min. d No significant clusters were formed before or after treatment with SERCA blocker TG (1 µM) in PTECs transfected with AMN-EYFP alone. Images were taken from Epi-Fluorescence (EPI-F) and TIRF microscopy. e Co-immunoprecipitation of STIM1, ORAI1 and AMN endogenously expressed in PTECs with/without TG treatment. f Colocalization of intracellular C terminus-truncated AMN (AMN1-183) with STIM1 in PTECs after Ca 2+ store depletion. Scale bars, 10 µm

    Journal: Nature Communications

    Article Title: ORAI channels are critical for receptor-mediated endocytosis of albumin

    doi: 10.1038/s41467-017-02094-y

    Figure Lengend Snippet: Subcellular localization and interaction of amnionless (AMN), STIM1 and ORAIs in PTECs. PTECs were cotransfected with AMN-EYFP, STIM1-mCherry and CFP-ORAIs and fluorescence was examined before and after treatment with thapsigargin (TG, 1 µM) for 10 min using TIRF microscopy with objective ( × 100). a Before TG treatment (Pre-TG). b After store-depletion with TG (post-TG). The fluorescence distribution pattern and intensity (below each image) are shown for cells transfected with CFP-ORAI1, CFP-ORAI2, and CFP-ORAI3, respectively, plus AMN-EYFP and STMI1-mCherry. c Pictures show the localization of STIM1-mCherry, CFP-ORAI1 and AMN-EYFP at the apical and basal membrane of human PTECs after treatment with 1 µM TG for 10 min. d No significant clusters were formed before or after treatment with SERCA blocker TG (1 µM) in PTECs transfected with AMN-EYFP alone. Images were taken from Epi-Fluorescence (EPI-F) and TIRF microscopy. e Co-immunoprecipitation of STIM1, ORAI1 and AMN endogenously expressed in PTECs with/without TG treatment. f Colocalization of intracellular C terminus-truncated AMN (AMN1-183) with STIM1 in PTECs after Ca 2+ store depletion. Scale bars, 10 µm

    Article Snippet: Immunoprecipitation of endogenously expressed STIM1, ORAI1 and AMN proteins in PTECs was performed with a Pierce Classic IP Kit (Thermo Fisher Scientific, USA), and rabbit anti-STIM1 at 1:500 (ACC-063, Alomone Labs, Jerusalem, Israel), rabbit anti-Orai1 at 1:500 (sc-68895, Santa Cruz) and mouse anti-AMN at 1:500 (sc-365384, Santa Cruz) antibodies were used.

    Techniques: Fluorescence, Microscopy, Transfection, Immunoprecipitation

    Model for ORAI/STIM and endocytic receptors in the process of albumin endocytosis Endoplasmic reticulum Ca 2+ store-depletion induces intracellular STIM1 movement and subplasmalemmal clustering, and forms ORAI/STIM1 complexes. The ORAI/STIM1 complexes physically associate with amnionless (AMN) to form triple complexes (ORAI/STIM1/AMN). Such triple complexes, together with cubilin and megalin, are endocytosed via clathrin-mediated endocytic mechanism during the PTECs exposure to albumin, which in turns leads to decreased activity of store-operated Ca 2+ entry in the cells

    Journal: Nature Communications

    Article Title: ORAI channels are critical for receptor-mediated endocytosis of albumin

    doi: 10.1038/s41467-017-02094-y

    Figure Lengend Snippet: Model for ORAI/STIM and endocytic receptors in the process of albumin endocytosis Endoplasmic reticulum Ca 2+ store-depletion induces intracellular STIM1 movement and subplasmalemmal clustering, and forms ORAI/STIM1 complexes. The ORAI/STIM1 complexes physically associate with amnionless (AMN) to form triple complexes (ORAI/STIM1/AMN). Such triple complexes, together with cubilin and megalin, are endocytosed via clathrin-mediated endocytic mechanism during the PTECs exposure to albumin, which in turns leads to decreased activity of store-operated Ca 2+ entry in the cells

    Article Snippet: Immunoprecipitation of endogenously expressed STIM1, ORAI1 and AMN proteins in PTECs was performed with a Pierce Classic IP Kit (Thermo Fisher Scientific, USA), and rabbit anti-STIM1 at 1:500 (ACC-063, Alomone Labs, Jerusalem, Israel), rabbit anti-Orai1 at 1:500 (sc-68895, Santa Cruz) and mouse anti-AMN at 1:500 (sc-365384, Santa Cruz) antibodies were used.

    Techniques: Activity Assay

    Clathrin-mediated endocytosis of albumin and ORAI/STIM complexes upon Ca 2+ store depletion. a , b , Localization of STIM1-EYFP, CFP-ORAI1, clathrin-mCherry and caveolin-1-mCherry at the apical and basal membrane of PTECs. c Percentage of STIM1 punctum areas that overlapped with ORAI1, clathrin or caveolin-1 ( n = 5). d Effects of endocytosis blockers chlorpromazine (50 µM) and nystatin (50 µM) on the uptake of FITC-albumin by PTECs ( n = 6 for each group). Average data are presented as mean ± s.e.m. The data sets are compared by ANOVA. Statistical significance is indicated by ** P

    Journal: Nature Communications

    Article Title: ORAI channels are critical for receptor-mediated endocytosis of albumin

    doi: 10.1038/s41467-017-02094-y

    Figure Lengend Snippet: Clathrin-mediated endocytosis of albumin and ORAI/STIM complexes upon Ca 2+ store depletion. a , b , Localization of STIM1-EYFP, CFP-ORAI1, clathrin-mCherry and caveolin-1-mCherry at the apical and basal membrane of PTECs. c Percentage of STIM1 punctum areas that overlapped with ORAI1, clathrin or caveolin-1 ( n = 5). d Effects of endocytosis blockers chlorpromazine (50 µM) and nystatin (50 µM) on the uptake of FITC-albumin by PTECs ( n = 6 for each group). Average data are presented as mean ± s.e.m. The data sets are compared by ANOVA. Statistical significance is indicated by ** P

    Article Snippet: Immunoprecipitation of endogenously expressed STIM1, ORAI1 and AMN proteins in PTECs was performed with a Pierce Classic IP Kit (Thermo Fisher Scientific, USA), and rabbit anti-STIM1 at 1:500 (ACC-063, Alomone Labs, Jerusalem, Israel), rabbit anti-Orai1 at 1:500 (sc-68895, Santa Cruz) and mouse anti-AMN at 1:500 (sc-365384, Santa Cruz) antibodies were used.

    Techniques:

    Subcellular localization of AMN, STIM1, and F-actin and the role of F-actin in albumin uptake and SOCE. a Images taken using Epi-Fluorescence (EPI-F) and TIRF before (pre-TG) and after 1 µM TG treatment for 10 min (post-TG). AMN tagged with EYFP (AMN-EYFP, green) and STIM1 tagged with mCherry (STIM1-Cherry, red). Subplasmalemmal distribution of F-actin was examined by transfecting PTECs with Lifeact-CFP to label F-actin. b Localization of STIM1-EYFP, mCherry-ORAI1 and F-actin at the apical and basal membranes after Ca 2+ store depletion. Scale bars, 10 µm. c Influence of cytochalasin D (CytD, 10 μM to depolymerize F-actin, n = 19), calyculin A (CalyA, 10 nM, to increase cortical F-actin content, n = 14) and U73122 (10 μM, to increase F-actin, n = 13) on store-operated Ca 2+ influx ( n = 12 in control group). Cells were pre-incubated with each drug for 30 min. d FITC-albumin uptake by PTECs was measured after pre-incubation with each drug for 30 min ( n = 6 for each group). Average data are presented as mean ± s.e.m. The data sets are compared by ANOVA. Statistical significance is indicated by ** P

    Journal: Nature Communications

    Article Title: ORAI channels are critical for receptor-mediated endocytosis of albumin

    doi: 10.1038/s41467-017-02094-y

    Figure Lengend Snippet: Subcellular localization of AMN, STIM1, and F-actin and the role of F-actin in albumin uptake and SOCE. a Images taken using Epi-Fluorescence (EPI-F) and TIRF before (pre-TG) and after 1 µM TG treatment for 10 min (post-TG). AMN tagged with EYFP (AMN-EYFP, green) and STIM1 tagged with mCherry (STIM1-Cherry, red). Subplasmalemmal distribution of F-actin was examined by transfecting PTECs with Lifeact-CFP to label F-actin. b Localization of STIM1-EYFP, mCherry-ORAI1 and F-actin at the apical and basal membranes after Ca 2+ store depletion. Scale bars, 10 µm. c Influence of cytochalasin D (CytD, 10 μM to depolymerize F-actin, n = 19), calyculin A (CalyA, 10 nM, to increase cortical F-actin content, n = 14) and U73122 (10 μM, to increase F-actin, n = 13) on store-operated Ca 2+ influx ( n = 12 in control group). Cells were pre-incubated with each drug for 30 min. d FITC-albumin uptake by PTECs was measured after pre-incubation with each drug for 30 min ( n = 6 for each group). Average data are presented as mean ± s.e.m. The data sets are compared by ANOVA. Statistical significance is indicated by ** P

    Article Snippet: Immunoprecipitation of endogenously expressed STIM1, ORAI1 and AMN proteins in PTECs was performed with a Pierce Classic IP Kit (Thermo Fisher Scientific, USA), and rabbit anti-STIM1 at 1:500 (ACC-063, Alomone Labs, Jerusalem, Israel), rabbit anti-Orai1 at 1:500 (sc-68895, Santa Cruz) and mouse anti-AMN at 1:500 (sc-365384, Santa Cruz) antibodies were used.

    Techniques: Fluorescence, Incubation

    ORAIs and STIMs in human kidney and regulation under diabetic condition. a Immunostaining for ORAIs in normal and diabetic kidney tissue sections. The diabetic kidney sections showing typical mesangial expansion and accumulation of mesangial matrix material. Scale bar, 100 µm. b , c Mean ± s.e.m. for the staining intensity (arbitrary unit) in proximal tubules and distal tubules, respectively. The average of nine staining fields for each patient was calculated for proximal or distal tubule staining ( n = 6 for normal kidney, n = 8 for diabetic kidney). Also see staining for STIM1 and STIM2 (Supplementary Fig. 3 ). d Primary cultured human proximal tubular epithelial cells (PTECs) were characterized by lectin staining (red). Scale bars, 50 µm. e PTECs were cultured with normal (5.5 mM) and high (25 mM) glucose for 60 h. ORAI proteins were detected by western blotting ( n = 6). f The mRNA of ORAIs was quantified by real-time PCR. The mean data were from 2–3 independent experiments ( n = 6). g The proximal tubular epithelial cells (HK-2) were treated with or without (control) insulin (10 nM) for 48 h and the mRNA was detected by real-time PCR ( n = 6). h HK-2 cells incubated with tyrosine kinase inhibitor tyrphostin A23 (30 µM) for 48 h ( n = 6). i STIM1 and STIM2 expression after insulin (10 nM) and tyrphostin A23 (30 µM) treatment for 48 h ( n = 9). The β-actin was used as control for relative quantification of mRNA or protein. For PCR experiments, triplicate reactions were set for each gene. The averaged data are displayed as mean ± s.e.m. and the data in e – i are normalized to control. The data sets are compared by t test. Statistical significance is indicated by * P

    Journal: Nature Communications

    Article Title: ORAI channels are critical for receptor-mediated endocytosis of albumin

    doi: 10.1038/s41467-017-02094-y

    Figure Lengend Snippet: ORAIs and STIMs in human kidney and regulation under diabetic condition. a Immunostaining for ORAIs in normal and diabetic kidney tissue sections. The diabetic kidney sections showing typical mesangial expansion and accumulation of mesangial matrix material. Scale bar, 100 µm. b , c Mean ± s.e.m. for the staining intensity (arbitrary unit) in proximal tubules and distal tubules, respectively. The average of nine staining fields for each patient was calculated for proximal or distal tubule staining ( n = 6 for normal kidney, n = 8 for diabetic kidney). Also see staining for STIM1 and STIM2 (Supplementary Fig. 3 ). d Primary cultured human proximal tubular epithelial cells (PTECs) were characterized by lectin staining (red). Scale bars, 50 µm. e PTECs were cultured with normal (5.5 mM) and high (25 mM) glucose for 60 h. ORAI proteins were detected by western blotting ( n = 6). f The mRNA of ORAIs was quantified by real-time PCR. The mean data were from 2–3 independent experiments ( n = 6). g The proximal tubular epithelial cells (HK-2) were treated with or without (control) insulin (10 nM) for 48 h and the mRNA was detected by real-time PCR ( n = 6). h HK-2 cells incubated with tyrosine kinase inhibitor tyrphostin A23 (30 µM) for 48 h ( n = 6). i STIM1 and STIM2 expression after insulin (10 nM) and tyrphostin A23 (30 µM) treatment for 48 h ( n = 9). The β-actin was used as control for relative quantification of mRNA or protein. For PCR experiments, triplicate reactions were set for each gene. The averaged data are displayed as mean ± s.e.m. and the data in e – i are normalized to control. The data sets are compared by t test. Statistical significance is indicated by * P

    Article Snippet: Immunoprecipitation of endogenously expressed STIM1, ORAI1 and AMN proteins in PTECs was performed with a Pierce Classic IP Kit (Thermo Fisher Scientific, USA), and rabbit anti-STIM1 at 1:500 (ACC-063, Alomone Labs, Jerusalem, Israel), rabbit anti-Orai1 at 1:500 (sc-68895, Santa Cruz) and mouse anti-AMN at 1:500 (sc-365384, Santa Cruz) antibodies were used.

    Techniques: Immunostaining, Staining, Cell Culture, Western Blot, Real-time Polymerase Chain Reaction, Incubation, Expressing, Polymerase Chain Reaction

    Schematic representation of the involvement of SERCA2a in the physiological control of SOCE GPCR - G-protein coupled receptor; PLC - phospholipase C; NFAT - nuclear factor of activated T lymphocytes; P - phosphate; IP 3 - inositol-1,4,5-trisphosphate, IP 3 R - IP 3 receptor; SR/ER sarco/endoplasmic reticulum; SERCA - SE/ER Ca 2+ ATPase; STIM1 - Stromal Interaction Molecule 1, ORAI1 - the pore forming unit.

    Journal: Journal of molecular and cellular cardiology

    Article Title: SERCA2a controls the mode of agonist-induced intracellular Ca2+ signal, transcription factor NFAT and proliferation in human vascular smooth muscle cells

    doi: 10.1016/j.yjmcc.2010.12.016

    Figure Lengend Snippet: Schematic representation of the involvement of SERCA2a in the physiological control of SOCE GPCR - G-protein coupled receptor; PLC - phospholipase C; NFAT - nuclear factor of activated T lymphocytes; P - phosphate; IP 3 - inositol-1,4,5-trisphosphate, IP 3 R - IP 3 receptor; SR/ER sarco/endoplasmic reticulum; SERCA - SE/ER Ca 2+ ATPase; STIM1 - Stromal Interaction Molecule 1, ORAI1 - the pore forming unit.

    Article Snippet: The following primary antibodies were used: IID8 (sc-53010, Santa Cruz Biotechnology), anti-SERCA2a and anti-SERCA2b [ ], anti-RyR2 [ ], anti- n on- m uscular myosin heavy chain B (NM-B) (Ab 684, Abcam), anti-smooth muscle m yosin h eavy c hain (MHC) (M3558, Dako Cytomation), anti-Cyclin D1 (556470, BD Biosciences), anti-PP2B (calcineurin, 556350, BD Biosciences), anti-STIM1 (ACC-63, Alomone labs), anti-Orai2 (ACC-061, Alomone labs), anti-ORAI1 (ACC-60, Alomone lab), anti-ORAI1 (sc-68895), anti-Cav1.2 calcium channel (L-type Ca2+ channel α1C subunit) (75053, NeuroMab); anti-h-calponin (C2687, Sigma-Aldrich), anti-caldesmon (C4562, Sigma-Aldrich); anti- g lycer a ldehyde 3- p hosphate d e h ydrogenase (GAPDH) (sc-47424, Santa Cruz Biotechnology).

    Techniques: Planar Chromatography

    SERCA2a prevents the formation of STIM-1/ORAI1 complex in cultured hCASMCs A. Effect of SERCA2a gene transfer on the expression of SOC sub-units. mRNA level quantified by real-time PCR was normalized to the value obtained in Ad-βGal-infected cells. Histograms show the means ± SEM of three experiments. B. Cells were infected for 4 days with Ad-βGal or Ad-S2a. Total protein extracts (50μg) were loaded. Left panel: western blot showing the expression of ORAI1, ORAI2 and STIM1 in whole-cell lysates. Right panel : histograms showing the relative ratio of ORAI1, ORAI2 and STIM1 normalized to GAPDH in three experiments. C. Whole-cell lysates were immunoprecipitated (IP) with an anti-STIM1 antibody, resolved on SDS/PAGE and immunobloted for ORAI1 or ORAI2. Membranes were reprobed for STIM1 for protein loading control. Histograms showing the mean (n=4) relative ratio of ORAI1 (left panel) and ORAI2 (right panel) normalized to STIM1 and arbitrary considered as 100% for Ad-βGal infected cells.

    Journal: Journal of molecular and cellular cardiology

    Article Title: SERCA2a controls the mode of agonist-induced intracellular Ca2+ signal, transcription factor NFAT and proliferation in human vascular smooth muscle cells

    doi: 10.1016/j.yjmcc.2010.12.016

    Figure Lengend Snippet: SERCA2a prevents the formation of STIM-1/ORAI1 complex in cultured hCASMCs A. Effect of SERCA2a gene transfer on the expression of SOC sub-units. mRNA level quantified by real-time PCR was normalized to the value obtained in Ad-βGal-infected cells. Histograms show the means ± SEM of three experiments. B. Cells were infected for 4 days with Ad-βGal or Ad-S2a. Total protein extracts (50μg) were loaded. Left panel: western blot showing the expression of ORAI1, ORAI2 and STIM1 in whole-cell lysates. Right panel : histograms showing the relative ratio of ORAI1, ORAI2 and STIM1 normalized to GAPDH in three experiments. C. Whole-cell lysates were immunoprecipitated (IP) with an anti-STIM1 antibody, resolved on SDS/PAGE and immunobloted for ORAI1 or ORAI2. Membranes were reprobed for STIM1 for protein loading control. Histograms showing the mean (n=4) relative ratio of ORAI1 (left panel) and ORAI2 (right panel) normalized to STIM1 and arbitrary considered as 100% for Ad-βGal infected cells.

    Article Snippet: The following primary antibodies were used: IID8 (sc-53010, Santa Cruz Biotechnology), anti-SERCA2a and anti-SERCA2b [ ], anti-RyR2 [ ], anti- n on- m uscular myosin heavy chain B (NM-B) (Ab 684, Abcam), anti-smooth muscle m yosin h eavy c hain (MHC) (M3558, Dako Cytomation), anti-Cyclin D1 (556470, BD Biosciences), anti-PP2B (calcineurin, 556350, BD Biosciences), anti-STIM1 (ACC-63, Alomone labs), anti-Orai2 (ACC-061, Alomone labs), anti-ORAI1 (ACC-60, Alomone lab), anti-ORAI1 (sc-68895), anti-Cav1.2 calcium channel (L-type Ca2+ channel α1C subunit) (75053, NeuroMab); anti-h-calponin (C2687, Sigma-Aldrich), anti-caldesmon (C4562, Sigma-Aldrich); anti- g lycer a ldehyde 3- p hosphate d e h ydrogenase (GAPDH) (sc-47424, Santa Cruz Biotechnology).

    Techniques: Cell Culture, Expressing, Real-time Polymerase Chain Reaction, Infection, Western Blot, Immunoprecipitation, SDS Page

    Increased TRPC1 and STIM1 in MetS are abolished by exercise. ( A ) Representative agarose gel image quantifying TRPC1 mRNA using RT–PCR. ( B ) TRPC1 mRNA normalized to β-actin. STIM1 ( C ) and Orai1 ( D ) mRNA using quantitative RT–PCR

    Journal: Cardiovascular Research

    Article Title: Exercise training decreases store-operated Ca2+entry associated with metabolic syndrome and coronary atherosclerosis

    doi: 10.1093/cvr/cvp308

    Figure Lengend Snippet: Increased TRPC1 and STIM1 in MetS are abolished by exercise. ( A ) Representative agarose gel image quantifying TRPC1 mRNA using RT–PCR. ( B ) TRPC1 mRNA normalized to β-actin. STIM1 ( C ) and Orai1 ( D ) mRNA using quantitative RT–PCR

    Article Snippet: STIM1 and ORAI1 antibodies were from Alomone labs (Rehovet, Israel).

    Techniques: Agarose Gel Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

    STIM1 deficiency promotes cardiomyocyte apoptosis induced by Dox. ( A – D ) Cardiomyocytes were treated with negative siRNA (Neg, 40 nmol/L), STIM1 siRNA (siSTIM1, 40 nmol/L) ( A ), Lacz adenovirus (Ad-Lacz, 40 MOI) or STIM1 adenovirus (Ad-STIM1, 40 MOI) ( B ) for 24 h in prior to Dox (10 μmol/L) incubation for another 24 h. Cell apoptosis was determined by Annexin V/PI staining followed by flow cytometry ( A and B ). Quantitative analysis of the percentage of apoptotic cells ( C and D ). ( E and F ) After the above-mentioned treatment, cell viability was assessed with CCK-8 assay. ( G and H ) At the end of the experiment, cell medium were harvested and LDH release was measured. Data were expressed as mean ± SEM. **P

    Journal: Journal of Inflammation Research

    Article Title: Cardiomyocyte Stim1 Deficiency Exacerbates Doxorubicin Cardiotoxicity by Magnification of Endoplasmic Reticulum Stress

    doi: 10.2147/JIR.S304520

    Figure Lengend Snippet: STIM1 deficiency promotes cardiomyocyte apoptosis induced by Dox. ( A – D ) Cardiomyocytes were treated with negative siRNA (Neg, 40 nmol/L), STIM1 siRNA (siSTIM1, 40 nmol/L) ( A ), Lacz adenovirus (Ad-Lacz, 40 MOI) or STIM1 adenovirus (Ad-STIM1, 40 MOI) ( B ) for 24 h in prior to Dox (10 μmol/L) incubation for another 24 h. Cell apoptosis was determined by Annexin V/PI staining followed by flow cytometry ( A and B ). Quantitative analysis of the percentage of apoptotic cells ( C and D ). ( E and F ) After the above-mentioned treatment, cell viability was assessed with CCK-8 assay. ( G and H ) At the end of the experiment, cell medium were harvested and LDH release was measured. Data were expressed as mean ± SEM. **P

    Article Snippet: Antibodies against STIM1, STIM2, ORAI1, and TRPC1 were obtained from Alomone (Jerusalem, Israel).

    Techniques: Incubation, Staining, Flow Cytometry, CCK-8 Assay

    Dox treatment reduces Stim1 expression and SOCE in cardiomyocytes. ( A ) Western blotting analysis of Stim1 expression in myocardium of mice after vehicle or Dox treatment for 14 days. **P

    Journal: Journal of Inflammation Research

    Article Title: Cardiomyocyte Stim1 Deficiency Exacerbates Doxorubicin Cardiotoxicity by Magnification of Endoplasmic Reticulum Stress

    doi: 10.2147/JIR.S304520

    Figure Lengend Snippet: Dox treatment reduces Stim1 expression and SOCE in cardiomyocytes. ( A ) Western blotting analysis of Stim1 expression in myocardium of mice after vehicle or Dox treatment for 14 days. **P

    Article Snippet: Antibodies against STIM1, STIM2, ORAI1, and TRPC1 were obtained from Alomone (Jerusalem, Israel).

    Techniques: Expressing, Western Blot, Mouse Assay

    Stim1 knockout exacerbates Dox-induced myocardial apoptosis. ( A and B ) Cell apoptosis in the cross-sectional area of myocardium from Stim1 CKO mice, Stim1 f/f mice ( A ), AAV-Lacz-infected mice and AAV-Stim1-infected mice ( B ) treated with vehicle or Dox was examined by TUNEL staining. Representative TUNEL (green) and α-actinin (red) stained photographs of cardiomyocytes in myocardial tissues. The nuclei was counterstained with DAPI (blue). n=6/group. Scale bar, 50 μm. ( C – J ) The protein expression of Bcl-2 and Bax in myocardium from the above groups were determined. Representative images of Western blotting were shown ( C and G ). Densitometric analysis of Bcl-2 ( D and H ), Bax ( E and I ), and the ratio of Bcl-2 to Bax ( F and J ) were performed. **P

    Journal: Journal of Inflammation Research

    Article Title: Cardiomyocyte Stim1 Deficiency Exacerbates Doxorubicin Cardiotoxicity by Magnification of Endoplasmic Reticulum Stress

    doi: 10.2147/JIR.S304520

    Figure Lengend Snippet: Stim1 knockout exacerbates Dox-induced myocardial apoptosis. ( A and B ) Cell apoptosis in the cross-sectional area of myocardium from Stim1 CKO mice, Stim1 f/f mice ( A ), AAV-Lacz-infected mice and AAV-Stim1-infected mice ( B ) treated with vehicle or Dox was examined by TUNEL staining. Representative TUNEL (green) and α-actinin (red) stained photographs of cardiomyocytes in myocardial tissues. The nuclei was counterstained with DAPI (blue). n=6/group. Scale bar, 50 μm. ( C – J ) The protein expression of Bcl-2 and Bax in myocardium from the above groups were determined. Representative images of Western blotting were shown ( C and G ). Densitometric analysis of Bcl-2 ( D and H ), Bax ( E and I ), and the ratio of Bcl-2 to Bax ( F and J ) were performed. **P

    Article Snippet: Antibodies against STIM1, STIM2, ORAI1, and TRPC1 were obtained from Alomone (Jerusalem, Israel).

    Techniques: Knock-Out, Mouse Assay, Infection, TUNEL Assay, Staining, Expressing, Western Blot

    Stim1 overexpression attenuates Dox-induced cardiotoxicity. ( A and B ) Mice were infected with AAV-Lacz or AAV-Stim1, and then infused with Dox (15 mg/kg of body weight) with an osmotic pump for 14 days. Serum LDH ( A ) and cTnT ( B ) concentration was examined. n=13/group. **P

    Journal: Journal of Inflammation Research

    Article Title: Cardiomyocyte Stim1 Deficiency Exacerbates Doxorubicin Cardiotoxicity by Magnification of Endoplasmic Reticulum Stress

    doi: 10.2147/JIR.S304520

    Figure Lengend Snippet: Stim1 overexpression attenuates Dox-induced cardiotoxicity. ( A and B ) Mice were infected with AAV-Lacz or AAV-Stim1, and then infused with Dox (15 mg/kg of body weight) with an osmotic pump for 14 days. Serum LDH ( A ) and cTnT ( B ) concentration was examined. n=13/group. **P

    Article Snippet: Antibodies against STIM1, STIM2, ORAI1, and TRPC1 were obtained from Alomone (Jerusalem, Israel).

    Techniques: Over Expression, Mouse Assay, Infection, Concentration Assay

    STIM1 relieves ER stress by binding to GRP78 in cardiomyocytes. ( A ) Representative images of STIM1 and GRP78 distribution in cardiomyocytes. ( B ) Cell lysates were immunoprecipitated with anti-GRP78 antibody and immunoprecipitated (IP) proteins were immunoblotted (IB) with anti-STIM1 antibody. n=5. ( C ) Cardiomyocytes were co-transfected with STIM1-flag plasmid and GRP78-myc plasmid. Anti-myc antibody was immunoprecipitated and immunoblotted with anti-flag antibody. n=4. ( D ) Immunoblotting for Stim1 after immunoprecipitation with anti-Grp78 antibody in myocardium from vehicle- or Dox-treated mice. **P

    Journal: Journal of Inflammation Research

    Article Title: Cardiomyocyte Stim1 Deficiency Exacerbates Doxorubicin Cardiotoxicity by Magnification of Endoplasmic Reticulum Stress

    doi: 10.2147/JIR.S304520

    Figure Lengend Snippet: STIM1 relieves ER stress by binding to GRP78 in cardiomyocytes. ( A ) Representative images of STIM1 and GRP78 distribution in cardiomyocytes. ( B ) Cell lysates were immunoprecipitated with anti-GRP78 antibody and immunoprecipitated (IP) proteins were immunoblotted (IB) with anti-STIM1 antibody. n=5. ( C ) Cardiomyocytes were co-transfected with STIM1-flag plasmid and GRP78-myc plasmid. Anti-myc antibody was immunoprecipitated and immunoblotted with anti-flag antibody. n=4. ( D ) Immunoblotting for Stim1 after immunoprecipitation with anti-Grp78 antibody in myocardium from vehicle- or Dox-treated mice. **P

    Article Snippet: Antibodies against STIM1, STIM2, ORAI1, and TRPC1 were obtained from Alomone (Jerusalem, Israel).

    Techniques: Binding Assay, Immunoprecipitation, Transfection, Plasmid Preparation, Mouse Assay

    Cardiomyocyte Stim1 deficiency enhances Dox-induced myocardial injury. ( A and B ) Cardiomyocyte-specific Stim1 knockout mice (Stim1 CKO ) and their control littermates (Stim1 f/f ) were treated with Dox (15 mg/kg body weight) for 14 days. LDH ( A ) and cTnT ( B ) release in serum were measured by a commercial kit. n=12/group. **P

    Journal: Journal of Inflammation Research

    Article Title: Cardiomyocyte Stim1 Deficiency Exacerbates Doxorubicin Cardiotoxicity by Magnification of Endoplasmic Reticulum Stress

    doi: 10.2147/JIR.S304520

    Figure Lengend Snippet: Cardiomyocyte Stim1 deficiency enhances Dox-induced myocardial injury. ( A and B ) Cardiomyocyte-specific Stim1 knockout mice (Stim1 CKO ) and their control littermates (Stim1 f/f ) were treated with Dox (15 mg/kg body weight) for 14 days. LDH ( A ) and cTnT ( B ) release in serum were measured by a commercial kit. n=12/group. **P

    Article Snippet: Antibodies against STIM1, STIM2, ORAI1, and TRPC1 were obtained from Alomone (Jerusalem, Israel).

    Techniques: Knock-Out, Mouse Assay

    STIM1 downregulation potentiates Dox-induced ER stress. ( A – D ) AC16 human cardiomyocytes were transfected with STIM1 siRNA (40 nmol/L) ( A and C ), STIM1 adenovirus (40 MOI) ( B and D ), or their corresponding negative control, and then incubated with Dox (10 μmol/L) for 1 h. The phosphorylation of PERK ( A and B ) and eiF2α ( C and D ) were determined. ( E – H ) The cells were treated with STIM1 siRNA ( E and G ) or STIM1 adenovirus ( F and H ) followed by incubation with Dox (10 μmol/L) for 24 h. The protein expression of ATF4 ( E and F ) and CHOP ( G and H ) were determined. Data were represented as mean ± SEM. **P

    Journal: Journal of Inflammation Research

    Article Title: Cardiomyocyte Stim1 Deficiency Exacerbates Doxorubicin Cardiotoxicity by Magnification of Endoplasmic Reticulum Stress

    doi: 10.2147/JIR.S304520

    Figure Lengend Snippet: STIM1 downregulation potentiates Dox-induced ER stress. ( A – D ) AC16 human cardiomyocytes were transfected with STIM1 siRNA (40 nmol/L) ( A and C ), STIM1 adenovirus (40 MOI) ( B and D ), or their corresponding negative control, and then incubated with Dox (10 μmol/L) for 1 h. The phosphorylation of PERK ( A and B ) and eiF2α ( C and D ) were determined. ( E – H ) The cells were treated with STIM1 siRNA ( E and G ) or STIM1 adenovirus ( F and H ) followed by incubation with Dox (10 μmol/L) for 24 h. The protein expression of ATF4 ( E and F ) and CHOP ( G and H ) were determined. Data were represented as mean ± SEM. **P

    Article Snippet: Antibodies against STIM1, STIM2, ORAI1, and TRPC1 were obtained from Alomone (Jerusalem, Israel).

    Techniques: Transfection, Negative Control, Incubation, Expressing