rabbit polyclonal ryr2  (Alomone Labs)


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    Alomone Labs rabbit polyclonal ryr2
    CCN5 prevents cardiac fibrosis in mdx/utrn (±) mice. (A) Experimental scheme for panels (B–E) . mdx/utrn (±) mice were injected with AAV.9-Con or CCN5 into the tail vein, and hearts were harvested for experiments 8 weeks later. Age-matched WT mice are shown in comparison. (B) Hearts were sectioned and stained with trichrome. Blue areas indicate fibrotic tissue and red areas indicate normal tissue. (C) The ratio of fibrotic area over total tissue of the stained hearts was plotted. (D) Proteins obtained from cardiac tissue were immunoblotted with antibodies against CCN5, α-SMA, collagen I, SERCA2a, <t>RyR2,</t> NCX1, phosphorylated phospholamban (p-PLN), t-PLN and α-tubulin. (E) Protein bands on western blots were scanned and plotted. n = 6. * p < 0.05 and ** p < 0.01.
    Rabbit Polyclonal Ryr2, 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 polyclonal ryr2/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal ryr2 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Matricellular Protein CCN5 Gene Transfer Ameliorates Cardiac and Skeletal Dysfunction in mdx/utrn (±) Haploinsufficient Mice by Reducing Fibrosis and Upregulating Utrophin Expression"

    Article Title: Matricellular Protein CCN5 Gene Transfer Ameliorates Cardiac and Skeletal Dysfunction in mdx/utrn (±) Haploinsufficient Mice by Reducing Fibrosis and Upregulating Utrophin Expression

    Journal: Frontiers in Cardiovascular Medicine

    doi: 10.3389/fcvm.2022.763544

    CCN5 prevents cardiac fibrosis in mdx/utrn (±) mice. (A) Experimental scheme for panels (B–E) . mdx/utrn (±) mice were injected with AAV.9-Con or CCN5 into the tail vein, and hearts were harvested for experiments 8 weeks later. Age-matched WT mice are shown in comparison. (B) Hearts were sectioned and stained with trichrome. Blue areas indicate fibrotic tissue and red areas indicate normal tissue. (C) The ratio of fibrotic area over total tissue of the stained hearts was plotted. (D) Proteins obtained from cardiac tissue were immunoblotted with antibodies against CCN5, α-SMA, collagen I, SERCA2a, RyR2, NCX1, phosphorylated phospholamban (p-PLN), t-PLN and α-tubulin. (E) Protein bands on western blots were scanned and plotted. n = 6. * p < 0.05 and ** p < 0.01.
    Figure Legend Snippet: CCN5 prevents cardiac fibrosis in mdx/utrn (±) mice. (A) Experimental scheme for panels (B–E) . mdx/utrn (±) mice were injected with AAV.9-Con or CCN5 into the tail vein, and hearts were harvested for experiments 8 weeks later. Age-matched WT mice are shown in comparison. (B) Hearts were sectioned and stained with trichrome. Blue areas indicate fibrotic tissue and red areas indicate normal tissue. (C) The ratio of fibrotic area over total tissue of the stained hearts was plotted. (D) Proteins obtained from cardiac tissue were immunoblotted with antibodies against CCN5, α-SMA, collagen I, SERCA2a, RyR2, NCX1, phosphorylated phospholamban (p-PLN), t-PLN and α-tubulin. (E) Protein bands on western blots were scanned and plotted. n = 6. * p < 0.05 and ** p < 0.01.

    Techniques Used: Injection, Staining, Western Blot

    anti pser2808 ryr2  (Alomone Labs)


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    Alomone Labs anti pser2808 ryr2
    Effects of macitentan on the calcium handling in LA in SHR. (a) Representative immunoblots showing expression of α1C, NCX1, CSQ and <t>RyR2</t> from SHR treated with MAC or DOX versus SHR-CTL (b-g) Quantification of immunoblots (N = 6–9 per group) (h) Representative immunoblots showing expression of SERCA2a, CaMKIIδ and PLB. Actin controls for pS16, pT17 and pS10 are not shown (i-m) Quantification of immunoblots (N = 6–9); # P < 0.05. Note that some blots/bands of housekeeping proteins used for normalization are identical (e.g. in (a) GAPDH blot/bands shown below CSQ and GAPDH blot/bands shown below RyR) as they are derived from the same membrane.
    Anti Pser2808 Ryr2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti pser2808 ryr2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti pser2808 ryr2 - by Bioz Stars, 2023-01
    86/100 stars

    Images

    1) Product Images from "Anti-inflammatory effects of endothelin receptor blockade in left atrial tissue of spontaneously hypertensive rats"

    Article Title: Anti-inflammatory effects of endothelin receptor blockade in left atrial tissue of spontaneously hypertensive rats

    Journal: International Journal of Cardiology. Heart & Vasculature

    doi: 10.1016/j.ijcha.2022.101088

    Effects of macitentan on the calcium handling in LA in SHR. (a) Representative immunoblots showing expression of α1C, NCX1, CSQ and RyR2 from SHR treated with MAC or DOX versus SHR-CTL (b-g) Quantification of immunoblots (N = 6–9 per group) (h) Representative immunoblots showing expression of SERCA2a, CaMKIIδ and PLB. Actin controls for pS16, pT17 and pS10 are not shown (i-m) Quantification of immunoblots (N = 6–9); # P < 0.05. Note that some blots/bands of housekeeping proteins used for normalization are identical (e.g. in (a) GAPDH blot/bands shown below CSQ and GAPDH blot/bands shown below RyR) as they are derived from the same membrane.
    Figure Legend Snippet: Effects of macitentan on the calcium handling in LA in SHR. (a) Representative immunoblots showing expression of α1C, NCX1, CSQ and RyR2 from SHR treated with MAC or DOX versus SHR-CTL (b-g) Quantification of immunoblots (N = 6–9 per group) (h) Representative immunoblots showing expression of SERCA2a, CaMKIIδ and PLB. Actin controls for pS16, pT17 and pS10 are not shown (i-m) Quantification of immunoblots (N = 6–9); # P < 0.05. Note that some blots/bands of housekeeping proteins used for normalization are identical (e.g. in (a) GAPDH blot/bands shown below CSQ and GAPDH blot/bands shown below RyR) as they are derived from the same membrane.

    Techniques Used: Western Blot, Expressing, Derivative Assay

    rabbit polyclonal ryr2  (Alomone Labs)


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    Alomone Labs rabbit polyclonal ryr2
    CCN5 prevents cardiac fibrosis in mdx/utrn (±) mice. (A) Experimental scheme for panels (B–E) . mdx/utrn (±) mice were injected with AAV.9-Con or CCN5 into the tail vein, and hearts were harvested for experiments 8 weeks later. Age-matched WT mice are shown in comparison. (B) Hearts were sectioned and stained with trichrome. Blue areas indicate fibrotic tissue and red areas indicate normal tissue. (C) The ratio of fibrotic area over total tissue of the stained hearts was plotted. (D) Proteins obtained from cardiac tissue were immunoblotted with antibodies against CCN5, α-SMA, collagen I, SERCA2a, <t>RyR2,</t> NCX1, phosphorylated phospholamban (p-PLN), t-PLN and α-tubulin. (E) Protein bands on western blots were scanned and plotted. n = 6. * p < 0.05 and ** p < 0.01.
    Rabbit Polyclonal Ryr2, 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 polyclonal ryr2/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit polyclonal ryr2 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Matricellular Protein CCN5 Gene Transfer Ameliorates Cardiac and Skeletal Dysfunction in mdx/utrn (±) Haploinsufficient Mice by Reducing Fibrosis and Upregulating Utrophin Expression"

    Article Title: Matricellular Protein CCN5 Gene Transfer Ameliorates Cardiac and Skeletal Dysfunction in mdx/utrn (±) Haploinsufficient Mice by Reducing Fibrosis and Upregulating Utrophin Expression

    Journal: Frontiers in Cardiovascular Medicine

    doi: 10.3389/fcvm.2022.763544

    CCN5 prevents cardiac fibrosis in mdx/utrn (±) mice. (A) Experimental scheme for panels (B–E) . mdx/utrn (±) mice were injected with AAV.9-Con or CCN5 into the tail vein, and hearts were harvested for experiments 8 weeks later. Age-matched WT mice are shown in comparison. (B) Hearts were sectioned and stained with trichrome. Blue areas indicate fibrotic tissue and red areas indicate normal tissue. (C) The ratio of fibrotic area over total tissue of the stained hearts was plotted. (D) Proteins obtained from cardiac tissue were immunoblotted with antibodies against CCN5, α-SMA, collagen I, SERCA2a, RyR2, NCX1, phosphorylated phospholamban (p-PLN), t-PLN and α-tubulin. (E) Protein bands on western blots were scanned and plotted. n = 6. * p < 0.05 and ** p < 0.01.
    Figure Legend Snippet: CCN5 prevents cardiac fibrosis in mdx/utrn (±) mice. (A) Experimental scheme for panels (B–E) . mdx/utrn (±) mice were injected with AAV.9-Con or CCN5 into the tail vein, and hearts were harvested for experiments 8 weeks later. Age-matched WT mice are shown in comparison. (B) Hearts were sectioned and stained with trichrome. Blue areas indicate fibrotic tissue and red areas indicate normal tissue. (C) The ratio of fibrotic area over total tissue of the stained hearts was plotted. (D) Proteins obtained from cardiac tissue were immunoblotted with antibodies against CCN5, α-SMA, collagen I, SERCA2a, RyR2, NCX1, phosphorylated phospholamban (p-PLN), t-PLN and α-tubulin. (E) Protein bands on western blots were scanned and plotted. n = 6. * p < 0.05 and ** p < 0.01.

    Techniques Used: Injection, Staining, Western Blot

    anti ryanodine receptor 2 ryr2  (Alomone Labs)


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    Alomone Labs anti ryanodine receptor 2 ryr2
    Discrete cAMP pools in adult mouse SAN cells (A) Diagrams highlighting localization and schematic representation of the Epac1-camps-based FRET biosensors (ICU3) in the cytosol (cyt; 1), plasma membrane (PM; 2), sarcoplasmic reticulum (SR; 3), myofilaments (MF; 4), and nucleus (nuc; 5). The ICU3 is linked to a Kras-derived sequence for PM localization, to a phospholamban (PLB)-derived sequence for SR localization, to a troponin T (TnT) for MF localization, and to a nuclear localization signal (NLS) sequence for nucleus localization. Exemplary super resolution images of adult wild-type mouse SAN cells expressing the indicated ICU3 biosensor in the cytosol (B), PM (C), SR (D), MF (E), and nucleus (F). The biosensor-associated fluorescence (YFP) is in magenta. Cells were immunostained with specific markers (in cyan) for the PM (caveolin 3), SR (ryanodine <t>receptor</t> <t>2</t> <t>[RyR2]),</t> MF (phalloidin; phal), and nucleus (DAPI). Merged images and corresponding line profile analysis (for dotted line) show high degree of overlap between the YFP fluorescence linked to the biosensor and the corresponding cellular marker in all cases, except in cells expressing the cytosolic sensor, as expected. Dotted squares highlight expanded regions in the solid squares. (G) Scatterplot of Pearson's correlation coefficient for cyt/cav3, PM/cav3, SR/RYR2, MF/phal, and nuc/DAPI (n > 8 SAN cells per condition). Kruskal-Wallis with Dunn's multiple comparisons test was used to test statistical differences in Pearson's correlation coefficient between non-target and targeted sensors. Scatterplot of the FRET ratio change in response to 10 μM forskolin (fsk) + 100 μM IBMX (H) and cAMP concentration-response curves (I) generated in HEK cells expressing the different ICU3 sensors (n > 5 cells per condition). For the cAMP concentration-response curves, cells expressing the different ICU3 sensors were exposed to increasing concentrations of the membrane-permeable cAMP analog 8CPT-cAMP. Kruskal-Wallis with Dunn's multiple comparisons test was used to compare fsk + IBMX responses, and the extra sum-of-squares F test was used to compare the cAMP EC 50 response between sensors. (J) Average FRET ratio traces (mean, solid lines; SEM, shadow) in response to 100 nM isoproterenol (iso) or 10 μM fsk from adult wild-type mouse SAN cells expressing the cytosolic, PM, SR, MF, or nuclear ICU3 biosensors (n > 5 cells from three preparations per condition). Scatterplots of ΔR/R 0 (K) and normalized (L) FRET responses after application of iso or fsk. Statistical differences were assessed with two-tailed Mann-Whitney test for comparisons between iso and fsk responses in (H) Statistical differences in fsk responses between the different biosensors in H were assessed with a Kruskal-Wallis with Dunn's multiple comparisons test. Statistical differences in normalized iso responses between the different groups were assessed using a one-way ANOVA with Tukey's multiple comparisons test. Significance (∗) was considered at P < 0.05. Exact p values are available in . Data represent mean ± SEM.
    Anti Ryanodine Receptor 2 Ryr2, 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/anti ryanodine receptor 2 ryr2/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti ryanodine receptor 2 ryr2 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Deciphering cellular signals in adult mouse sinoatrial node cells"

    Article Title: Deciphering cellular signals in adult mouse sinoatrial node cells

    Journal: iScience

    doi: 10.1016/j.isci.2021.103693

    Discrete cAMP pools in adult mouse SAN cells (A) Diagrams highlighting localization and schematic representation of the Epac1-camps-based FRET biosensors (ICU3) in the cytosol (cyt; 1), plasma membrane (PM; 2), sarcoplasmic reticulum (SR; 3), myofilaments (MF; 4), and nucleus (nuc; 5). The ICU3 is linked to a Kras-derived sequence for PM localization, to a phospholamban (PLB)-derived sequence for SR localization, to a troponin T (TnT) for MF localization, and to a nuclear localization signal (NLS) sequence for nucleus localization. Exemplary super resolution images of adult wild-type mouse SAN cells expressing the indicated ICU3 biosensor in the cytosol (B), PM (C), SR (D), MF (E), and nucleus (F). The biosensor-associated fluorescence (YFP) is in magenta. Cells were immunostained with specific markers (in cyan) for the PM (caveolin 3), SR (ryanodine receptor 2 [RyR2]), MF (phalloidin; phal), and nucleus (DAPI). Merged images and corresponding line profile analysis (for dotted line) show high degree of overlap between the YFP fluorescence linked to the biosensor and the corresponding cellular marker in all cases, except in cells expressing the cytosolic sensor, as expected. Dotted squares highlight expanded regions in the solid squares. (G) Scatterplot of Pearson's correlation coefficient for cyt/cav3, PM/cav3, SR/RYR2, MF/phal, and nuc/DAPI (n > 8 SAN cells per condition). Kruskal-Wallis with Dunn's multiple comparisons test was used to test statistical differences in Pearson's correlation coefficient between non-target and targeted sensors. Scatterplot of the FRET ratio change in response to 10 μM forskolin (fsk) + 100 μM IBMX (H) and cAMP concentration-response curves (I) generated in HEK cells expressing the different ICU3 sensors (n > 5 cells per condition). For the cAMP concentration-response curves, cells expressing the different ICU3 sensors were exposed to increasing concentrations of the membrane-permeable cAMP analog 8CPT-cAMP. Kruskal-Wallis with Dunn's multiple comparisons test was used to compare fsk + IBMX responses, and the extra sum-of-squares F test was used to compare the cAMP EC 50 response between sensors. (J) Average FRET ratio traces (mean, solid lines; SEM, shadow) in response to 100 nM isoproterenol (iso) or 10 μM fsk from adult wild-type mouse SAN cells expressing the cytosolic, PM, SR, MF, or nuclear ICU3 biosensors (n > 5 cells from three preparations per condition). Scatterplots of ΔR/R 0 (K) and normalized (L) FRET responses after application of iso or fsk. Statistical differences were assessed with two-tailed Mann-Whitney test for comparisons between iso and fsk responses in (H) Statistical differences in fsk responses between the different biosensors in H were assessed with a Kruskal-Wallis with Dunn's multiple comparisons test. Statistical differences in normalized iso responses between the different groups were assessed using a one-way ANOVA with Tukey's multiple comparisons test. Significance (∗) was considered at P < 0.05. Exact p values are available in . Data represent mean ± SEM.
    Figure Legend Snippet: Discrete cAMP pools in adult mouse SAN cells (A) Diagrams highlighting localization and schematic representation of the Epac1-camps-based FRET biosensors (ICU3) in the cytosol (cyt; 1), plasma membrane (PM; 2), sarcoplasmic reticulum (SR; 3), myofilaments (MF; 4), and nucleus (nuc; 5). The ICU3 is linked to a Kras-derived sequence for PM localization, to a phospholamban (PLB)-derived sequence for SR localization, to a troponin T (TnT) for MF localization, and to a nuclear localization signal (NLS) sequence for nucleus localization. Exemplary super resolution images of adult wild-type mouse SAN cells expressing the indicated ICU3 biosensor in the cytosol (B), PM (C), SR (D), MF (E), and nucleus (F). The biosensor-associated fluorescence (YFP) is in magenta. Cells were immunostained with specific markers (in cyan) for the PM (caveolin 3), SR (ryanodine receptor 2 [RyR2]), MF (phalloidin; phal), and nucleus (DAPI). Merged images and corresponding line profile analysis (for dotted line) show high degree of overlap between the YFP fluorescence linked to the biosensor and the corresponding cellular marker in all cases, except in cells expressing the cytosolic sensor, as expected. Dotted squares highlight expanded regions in the solid squares. (G) Scatterplot of Pearson's correlation coefficient for cyt/cav3, PM/cav3, SR/RYR2, MF/phal, and nuc/DAPI (n > 8 SAN cells per condition). Kruskal-Wallis with Dunn's multiple comparisons test was used to test statistical differences in Pearson's correlation coefficient between non-target and targeted sensors. Scatterplot of the FRET ratio change in response to 10 μM forskolin (fsk) + 100 μM IBMX (H) and cAMP concentration-response curves (I) generated in HEK cells expressing the different ICU3 sensors (n > 5 cells per condition). For the cAMP concentration-response curves, cells expressing the different ICU3 sensors were exposed to increasing concentrations of the membrane-permeable cAMP analog 8CPT-cAMP. Kruskal-Wallis with Dunn's multiple comparisons test was used to compare fsk + IBMX responses, and the extra sum-of-squares F test was used to compare the cAMP EC 50 response between sensors. (J) Average FRET ratio traces (mean, solid lines; SEM, shadow) in response to 100 nM isoproterenol (iso) or 10 μM fsk from adult wild-type mouse SAN cells expressing the cytosolic, PM, SR, MF, or nuclear ICU3 biosensors (n > 5 cells from three preparations per condition). Scatterplots of ΔR/R 0 (K) and normalized (L) FRET responses after application of iso or fsk. Statistical differences were assessed with two-tailed Mann-Whitney test for comparisons between iso and fsk responses in (H) Statistical differences in fsk responses between the different biosensors in H were assessed with a Kruskal-Wallis with Dunn's multiple comparisons test. Statistical differences in normalized iso responses between the different groups were assessed using a one-way ANOVA with Tukey's multiple comparisons test. Significance (∗) was considered at P < 0.05. Exact p values are available in . Data represent mean ± SEM.

    Techniques Used: Derivative Assay, Sequencing, Expressing, Fluorescence, Marker, Concentration Assay, Generated, Two Tailed Test, MANN-WHITNEY


    Figure Legend Snippet:

    Techniques Used: Recombinant, Software

    ryr2  (Alomone Labs)


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    Alomone Labs ryr2
    Ca2+ handling protein expression or phosphorylation at ZT4 and ZT14 in adult hearts. A-B. Phosphorylation of PLB at serine16 was essentially non-detectable following 5min perfusion (A) and 1hr perfusion (to match optical mapping data collection timepoint, B). Band labeled ‘Iso’ in (B) was a single heart perfused with isoproterenol for a positive control for pPLB. C-D. PLB phosphorylation was higher at the Thr17 site at ZT4 vs. ZT14 following 5min perfusion (C). However, phosphorylation levels were reduced and were not different between ZT4 and ZT14 following 1hr of perfusion (D). Band labeled ‘Iso’ in (D) was a single heart perfused with isoproterenol for a positive control for pPLB. E-G. Total PLB (E), SERCA2a (F), and <t>RyR2</t> (G) expression. α-tubulin was used as loading control. ZT4 n=4; ZT14 n=4; except RyR (G) where ZT4 n=3; ZT14 n=3. *p<0.05.
    Ryr2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ryr2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ryr2 - by Bioz Stars, 2023-01
    86/100 stars

    Images

    1) Product Images from "Aging Disrupts Normal Time-of-day Variation in Cardiac Electrophysiology"

    Article Title: Aging Disrupts Normal Time-of-day Variation in Cardiac Electrophysiology

    Journal: Circulation. Arrhythmia and electrophysiology

    doi: 10.1161/CIRCEP.119.008093

    Ca2+ handling protein expression or phosphorylation at ZT4 and ZT14 in adult hearts. A-B. Phosphorylation of PLB at serine16 was essentially non-detectable following 5min perfusion (A) and 1hr perfusion (to match optical mapping data collection timepoint, B). Band labeled ‘Iso’ in (B) was a single heart perfused with isoproterenol for a positive control for pPLB. C-D. PLB phosphorylation was higher at the Thr17 site at ZT4 vs. ZT14 following 5min perfusion (C). However, phosphorylation levels were reduced and were not different between ZT4 and ZT14 following 1hr of perfusion (D). Band labeled ‘Iso’ in (D) was a single heart perfused with isoproterenol for a positive control for pPLB. E-G. Total PLB (E), SERCA2a (F), and RyR2 (G) expression. α-tubulin was used as loading control. ZT4 n=4; ZT14 n=4; except RyR (G) where ZT4 n=3; ZT14 n=3. *p<0.05.
    Figure Legend Snippet: Ca2+ handling protein expression or phosphorylation at ZT4 and ZT14 in adult hearts. A-B. Phosphorylation of PLB at serine16 was essentially non-detectable following 5min perfusion (A) and 1hr perfusion (to match optical mapping data collection timepoint, B). Band labeled ‘Iso’ in (B) was a single heart perfused with isoproterenol for a positive control for pPLB. C-D. PLB phosphorylation was higher at the Thr17 site at ZT4 vs. ZT14 following 5min perfusion (C). However, phosphorylation levels were reduced and were not different between ZT4 and ZT14 following 1hr of perfusion (D). Band labeled ‘Iso’ in (D) was a single heart perfused with isoproterenol for a positive control for pPLB. E-G. Total PLB (E), SERCA2a (F), and RyR2 (G) expression. α-tubulin was used as loading control. ZT4 n=4; ZT14 n=4; except RyR (G) where ZT4 n=3; ZT14 n=3. *p<0.05.

    Techniques Used: Expressing, Labeling, Positive Control

    ryr2  (Alomone Labs)


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    Alomone Labs ryr2
    Ca2+ handling protein expression or phosphorylation at ZT4 and ZT14 in adult hearts. A-B. Phosphorylation of PLB at serine16 was essentially non-detectable following 5min perfusion (A) and 1hr perfusion (to match optical mapping data collection timepoint, B). Band labeled ‘Iso’ in (B) was a single heart perfused with isoproterenol for a positive control for pPLB. C-D. PLB phosphorylation was higher at the Thr17 site at ZT4 vs. ZT14 following 5min perfusion (C). However, phosphorylation levels were reduced and were not different between ZT4 and ZT14 following 1hr of perfusion (D). Band labeled ‘Iso’ in (D) was a single heart perfused with isoproterenol for a positive control for pPLB. E-G. Total PLB (E), SERCA2a (F), and <t>RyR2</t> (G) expression. α-tubulin was used as loading control. ZT4 n=4; ZT14 n=4; except RyR (G) where ZT4 n=3; ZT14 n=3. *p<0.05.
    Ryr2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ryr2/product/Alomone Labs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ryr2 - by Bioz Stars, 2023-01
    86/100 stars

    Images

    1) Product Images from "Aging Disrupts Normal Time-of-day Variation in Cardiac Electrophysiology"

    Article Title: Aging Disrupts Normal Time-of-day Variation in Cardiac Electrophysiology

    Journal: Circulation. Arrhythmia and electrophysiology

    doi: 10.1161/CIRCEP.119.008093

    Ca2+ handling protein expression or phosphorylation at ZT4 and ZT14 in adult hearts. A-B. Phosphorylation of PLB at serine16 was essentially non-detectable following 5min perfusion (A) and 1hr perfusion (to match optical mapping data collection timepoint, B). Band labeled ‘Iso’ in (B) was a single heart perfused with isoproterenol for a positive control for pPLB. C-D. PLB phosphorylation was higher at the Thr17 site at ZT4 vs. ZT14 following 5min perfusion (C). However, phosphorylation levels were reduced and were not different between ZT4 and ZT14 following 1hr of perfusion (D). Band labeled ‘Iso’ in (D) was a single heart perfused with isoproterenol for a positive control for pPLB. E-G. Total PLB (E), SERCA2a (F), and RyR2 (G) expression. α-tubulin was used as loading control. ZT4 n=4; ZT14 n=4; except RyR (G) where ZT4 n=3; ZT14 n=3. *p<0.05.
    Figure Legend Snippet: Ca2+ handling protein expression or phosphorylation at ZT4 and ZT14 in adult hearts. A-B. Phosphorylation of PLB at serine16 was essentially non-detectable following 5min perfusion (A) and 1hr perfusion (to match optical mapping data collection timepoint, B). Band labeled ‘Iso’ in (B) was a single heart perfused with isoproterenol for a positive control for pPLB. C-D. PLB phosphorylation was higher at the Thr17 site at ZT4 vs. ZT14 following 5min perfusion (C). However, phosphorylation levels were reduced and were not different between ZT4 and ZT14 following 1hr of perfusion (D). Band labeled ‘Iso’ in (D) was a single heart perfused with isoproterenol for a positive control for pPLB. E-G. Total PLB (E), SERCA2a (F), and RyR2 (G) expression. α-tubulin was used as loading control. ZT4 n=4; ZT14 n=4; except RyR (G) where ZT4 n=3; ZT14 n=3. *p<0.05.

    Techniques Used: Expressing, Labeling, Positive Control

    antibody ryr1  (Alomone Labs)


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    Alomone Labs antibody ryr1
    Flow cytometry detects <t>RYR1</t> in freshly isolated LMCs. Scatter plots of LMC suspensions plotted as side scatter (SSC-W) vs. fluorescent signals. Black horizontal lines denote the gate set. Fluorescent signal appearing above gate indicates positive staining. (A) Negative control (Neg) showing background FITC fluorescence of cell population with no antibody staining. Another LMC suspension was used to identify (B, left ) the LMC population stained by α-SMA-FITC ( red box ) and (B, right ) the gate set for non-specific staining of Alexa-Fluor 647 conjugated secondary antibody without primary antibodies within the α-SMA-FITC gated region. (C) Positive detection of RYR1 above the Alexa-Fluor 674 gate in LMCs isolated from rat mesenteric LVs. Data representative of five isolations each from male and female rats ( n = 10).
    Antibody Ryr1, 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
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    Average 93 stars, based on 1 article reviews
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    93/100 stars

    Images

    1) Product Images from "Dantrolene Prevents the Lymphostasis Caused by Doxorubicin in the Rat Mesenteric Circulation"

    Article Title: Dantrolene Prevents the Lymphostasis Caused by Doxorubicin in the Rat Mesenteric Circulation

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2021.727526

    Flow cytometry detects RYR1 in freshly isolated LMCs. Scatter plots of LMC suspensions plotted as side scatter (SSC-W) vs. fluorescent signals. Black horizontal lines denote the gate set. Fluorescent signal appearing above gate indicates positive staining. (A) Negative control (Neg) showing background FITC fluorescence of cell population with no antibody staining. Another LMC suspension was used to identify (B, left ) the LMC population stained by α-SMA-FITC ( red box ) and (B, right ) the gate set for non-specific staining of Alexa-Fluor 647 conjugated secondary antibody without primary antibodies within the α-SMA-FITC gated region. (C) Positive detection of RYR1 above the Alexa-Fluor 674 gate in LMCs isolated from rat mesenteric LVs. Data representative of five isolations each from male and female rats ( n = 10).
    Figure Legend Snippet: Flow cytometry detects RYR1 in freshly isolated LMCs. Scatter plots of LMC suspensions plotted as side scatter (SSC-W) vs. fluorescent signals. Black horizontal lines denote the gate set. Fluorescent signal appearing above gate indicates positive staining. (A) Negative control (Neg) showing background FITC fluorescence of cell population with no antibody staining. Another LMC suspension was used to identify (B, left ) the LMC population stained by α-SMA-FITC ( red box ) and (B, right ) the gate set for non-specific staining of Alexa-Fluor 647 conjugated secondary antibody without primary antibodies within the α-SMA-FITC gated region. (C) Positive detection of RYR1 above the Alexa-Fluor 674 gate in LMCs isolated from rat mesenteric LVs. Data representative of five isolations each from male and female rats ( n = 10).

    Techniques Used: Flow Cytometry, Isolation, Staining, Negative Control, Fluorescence

    RYR1 antibody selectivity. (A) Western blot shows positive detection of RYR1 (∼565 kD) in rat skeletal muscle lysate and no detection in rat heart or brain lysate. (B,C) Contour plots of RYR1 antibody testing. (B) Red population represents positive detection of RYR1 subtype in male LMCs. Grey population represents the LMC suspension containing no primary antibody. (C) No detection in male LMCs after co-incubation with competing peptide (CP; 1:50 dilution) for the RYR1 antibody. Data representative of three isolations for each sex ( n = 6).
    Figure Legend Snippet: RYR1 antibody selectivity. (A) Western blot shows positive detection of RYR1 (∼565 kD) in rat skeletal muscle lysate and no detection in rat heart or brain lysate. (B,C) Contour plots of RYR1 antibody testing. (B) Red population represents positive detection of RYR1 subtype in male LMCs. Grey population represents the LMC suspension containing no primary antibody. (C) No detection in male LMCs after co-incubation with competing peptide (CP; 1:50 dilution) for the RYR1 antibody. Data representative of three isolations for each sex ( n = 6).

    Techniques Used: Western Blot, Incubation

    ryr1 subtype  (Alomone Labs)


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    Alomone Labs ryr1 subtype
    Flow cytometry detects <t>RYR1</t> in freshly isolated LMCs. Scatter plots of LMC suspensions plotted as side scatter (SSC-W) vs. fluorescent signals. Black horizontal lines denote the gate set. Fluorescent signal appearing above gate indicates positive staining. (A) Negative control (Neg) showing background FITC fluorescence of cell population with no antibody staining. Another LMC suspension was used to identify (B, left ) the LMC population stained by α-SMA-FITC ( red box ) and (B, right ) the gate set for non-specific staining of Alexa-Fluor 647 conjugated secondary antibody without primary antibodies within the α-SMA-FITC gated region. (C) Positive detection of RYR1 above the Alexa-Fluor 674 gate in LMCs isolated from rat mesenteric LVs. Data representative of five isolations each from male and female rats ( n = 10).
    Ryr1 Subtype, 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
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    Average 93 stars, based on 1 article reviews
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    ryr1 subtype - by Bioz Stars, 2023-01
    93/100 stars

    Images

    1) Product Images from "Dantrolene Prevents the Lymphostasis Caused by Doxorubicin in the Rat Mesenteric Circulation"

    Article Title: Dantrolene Prevents the Lymphostasis Caused by Doxorubicin in the Rat Mesenteric Circulation

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2021.727526

    Flow cytometry detects RYR1 in freshly isolated LMCs. Scatter plots of LMC suspensions plotted as side scatter (SSC-W) vs. fluorescent signals. Black horizontal lines denote the gate set. Fluorescent signal appearing above gate indicates positive staining. (A) Negative control (Neg) showing background FITC fluorescence of cell population with no antibody staining. Another LMC suspension was used to identify (B, left ) the LMC population stained by α-SMA-FITC ( red box ) and (B, right ) the gate set for non-specific staining of Alexa-Fluor 647 conjugated secondary antibody without primary antibodies within the α-SMA-FITC gated region. (C) Positive detection of RYR1 above the Alexa-Fluor 674 gate in LMCs isolated from rat mesenteric LVs. Data representative of five isolations each from male and female rats ( n = 10).
    Figure Legend Snippet: Flow cytometry detects RYR1 in freshly isolated LMCs. Scatter plots of LMC suspensions plotted as side scatter (SSC-W) vs. fluorescent signals. Black horizontal lines denote the gate set. Fluorescent signal appearing above gate indicates positive staining. (A) Negative control (Neg) showing background FITC fluorescence of cell population with no antibody staining. Another LMC suspension was used to identify (B, left ) the LMC population stained by α-SMA-FITC ( red box ) and (B, right ) the gate set for non-specific staining of Alexa-Fluor 647 conjugated secondary antibody without primary antibodies within the α-SMA-FITC gated region. (C) Positive detection of RYR1 above the Alexa-Fluor 674 gate in LMCs isolated from rat mesenteric LVs. Data representative of five isolations each from male and female rats ( n = 10).

    Techniques Used: Flow Cytometry, Isolation, Staining, Negative Control, Fluorescence

    RYR1 antibody selectivity. (A) Western blot shows positive detection of RYR1 (∼565 kD) in rat skeletal muscle lysate and no detection in rat heart or brain lysate. (B,C) Contour plots of RYR1 antibody testing. (B) Red population represents positive detection of RYR1 subtype in male LMCs. Grey population represents the LMC suspension containing no primary antibody. (C) No detection in male LMCs after co-incubation with competing peptide (CP; 1:50 dilution) for the RYR1 antibody. Data representative of three isolations for each sex ( n = 6).
    Figure Legend Snippet: RYR1 antibody selectivity. (A) Western blot shows positive detection of RYR1 (∼565 kD) in rat skeletal muscle lysate and no detection in rat heart or brain lysate. (B,C) Contour plots of RYR1 antibody testing. (B) Red population represents positive detection of RYR1 subtype in male LMCs. Grey population represents the LMC suspension containing no primary antibody. (C) No detection in male LMCs after co-incubation with competing peptide (CP; 1:50 dilution) for the RYR1 antibody. Data representative of three isolations for each sex ( n = 6).

    Techniques Used: Western Blot, Incubation

    anti p s2814  (Alomone Labs)


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    Structured Review

    Alomone Labs anti p s2814
    Knockdown of TRPC7 decreased the activity of <t>RyR2</t> and SERCA without affecting the activity of IP3R in mESC-CMs. Representative CaTs recorded from the control group a before and b after applying ryanodine. Representative CaTs recorded from TRPC7 knockdown group c before and d after applying ryanodine. e , f Bar charts showing the normalized frequency and the V max-upstroke calculated from a – d . The decrease of the frequency and the V max-upstroke caused by ryanodine was less in TRPC7 knockdown group. Representative CaTs recorded from the control group g before and h after applying thapsigargin (TG). Representative CaTs recorded from TRPC7 knockdown group i before and j after applying TG. k , l Bar charts showing the normalized frequency and V max-decay calculated from g – j . The decrease of the frequency and the V max-decay and amplitude caused by TG was less in TRPC7 knockdown group. Representative CaTs recorded from the control group m before and n after applying 2-aminoethoxydipheylborate (2-APB). Representative CaTs recorded from TRPC7 knockdown group o before and p after applying 2-APB. q , r Bar charts showing the normalized frequency and the V max-upstroke calculated from m – p . The decrease of the frequency and the V max-upstroke caused by 2-APB was similar between control and TRPC7 knockdown groups. Data were presented as mean ± SEM ( n = 10 cells; cells were from 3 independent batches of differentiation). * P < 0.05, ** P < 0.01 vs control
    Anti P S2814, 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
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    anti p s2814 - by Bioz Stars, 2023-01
    93/100 stars

    Images

    1) Product Images from "TRPC7 regulates the electrophysiological functions of embryonic stem cell-derived cardiomyocytes"

    Article Title: TRPC7 regulates the electrophysiological functions of embryonic stem cell-derived cardiomyocytes

    Journal: Stem Cell Research & Therapy

    doi: 10.1186/s13287-021-02308-7

    Knockdown of TRPC7 decreased the activity of RyR2 and SERCA without affecting the activity of IP3R in mESC-CMs. Representative CaTs recorded from the control group a before and b after applying ryanodine. Representative CaTs recorded from TRPC7 knockdown group c before and d after applying ryanodine. e , f Bar charts showing the normalized frequency and the V max-upstroke calculated from a – d . The decrease of the frequency and the V max-upstroke caused by ryanodine was less in TRPC7 knockdown group. Representative CaTs recorded from the control group g before and h after applying thapsigargin (TG). Representative CaTs recorded from TRPC7 knockdown group i before and j after applying TG. k , l Bar charts showing the normalized frequency and V max-decay calculated from g – j . The decrease of the frequency and the V max-decay and amplitude caused by TG was less in TRPC7 knockdown group. Representative CaTs recorded from the control group m before and n after applying 2-aminoethoxydipheylborate (2-APB). Representative CaTs recorded from TRPC7 knockdown group o before and p after applying 2-APB. q , r Bar charts showing the normalized frequency and the V max-upstroke calculated from m – p . The decrease of the frequency and the V max-upstroke caused by 2-APB was similar between control and TRPC7 knockdown groups. Data were presented as mean ± SEM ( n = 10 cells; cells were from 3 independent batches of differentiation). * P < 0.05, ** P < 0.01 vs control
    Figure Legend Snippet: Knockdown of TRPC7 decreased the activity of RyR2 and SERCA without affecting the activity of IP3R in mESC-CMs. Representative CaTs recorded from the control group a before and b after applying ryanodine. Representative CaTs recorded from TRPC7 knockdown group c before and d after applying ryanodine. e , f Bar charts showing the normalized frequency and the V max-upstroke calculated from a – d . The decrease of the frequency and the V max-upstroke caused by ryanodine was less in TRPC7 knockdown group. Representative CaTs recorded from the control group g before and h after applying thapsigargin (TG). Representative CaTs recorded from TRPC7 knockdown group i before and j after applying TG. k , l Bar charts showing the normalized frequency and V max-decay calculated from g – j . The decrease of the frequency and the V max-decay and amplitude caused by TG was less in TRPC7 knockdown group. Representative CaTs recorded from the control group m before and n after applying 2-aminoethoxydipheylborate (2-APB). Representative CaTs recorded from TRPC7 knockdown group o before and p after applying 2-APB. q , r Bar charts showing the normalized frequency and the V max-upstroke calculated from m – p . The decrease of the frequency and the V max-upstroke caused by 2-APB was similar between control and TRPC7 knockdown groups. Data were presented as mean ± SEM ( n = 10 cells; cells were from 3 independent batches of differentiation). * P < 0.05, ** P < 0.01 vs control

    Techniques Used: Activity Assay

    Knockdown of TRPC7 decreased while overexpression of TRPC7 increased the phosphorylation of RyR2 and PLN, respectively, in NRVMs. Western blotting showing the expression of a total RyR2, b p(S2814)RyR2, c total PLN, and d p(T17) PLN in NRVMs infected with different adenoviruses to knockdown or overexpress TRPC7. e – h Bar charts showing the quantification of each protein from a – d . To eliminate the loading bias, intensity of each target protein was normalized to that of its corresponding β-tubulin. The change of TRPC7 expression did not alter the expression of total RyR2 and PLN. However, knockdown of TRPC7 decreased while overexpression of TRPC7 increased the phosphorylation form of RyR2 [p(S2814)RyR2] and PLN [p(T17)PLN]. Data were presented as mean ± SEM ( n = 4). * P < 0.05, *** P < 0.001 vs corresponding controls
    Figure Legend Snippet: Knockdown of TRPC7 decreased while overexpression of TRPC7 increased the phosphorylation of RyR2 and PLN, respectively, in NRVMs. Western blotting showing the expression of a total RyR2, b p(S2814)RyR2, c total PLN, and d p(T17) PLN in NRVMs infected with different adenoviruses to knockdown or overexpress TRPC7. e – h Bar charts showing the quantification of each protein from a – d . To eliminate the loading bias, intensity of each target protein was normalized to that of its corresponding β-tubulin. The change of TRPC7 expression did not alter the expression of total RyR2 and PLN. However, knockdown of TRPC7 decreased while overexpression of TRPC7 increased the phosphorylation form of RyR2 [p(S2814)RyR2] and PLN [p(T17)PLN]. Data were presented as mean ± SEM ( n = 4). * P < 0.05, *** P < 0.001 vs corresponding controls

    Techniques Used: Over Expression, Western Blot, Expressing, Infection

    Schematic diagram illustrating the mechanism through which TRPC7 positively regulates the automaticity of cardiomyocytes. G protein-coupled receptors (GPCRs) locating on the plasma membrane (PM) sense the external ligands such as hormones, neurotransmitters, and growth factors, transduce the signal to activate phospholipase C (PLC) which hydrolyzes the phosphatidylinositol 4,5-bisphosphate (PIP 2 ) into inositol trisphosphate (IP 3 ) and diacylglycerol (DAG). TRPC7 is then directly activated by DAG, mediating the Ca 2+ influx. Ca 2+ permeated through TRPC7 may activate the forward mode of the Na + -Ca 2+ exchanger (NCX), leading Na + influx and depolarization. This depolarization would then accelerate diastolic depolarization (DD) and increase AP firing rate. On the other hand, Ca 2+ permeated through TRPC7 may also increase the activity of ryanodine receptor 2 (RyR2) locating on the SR, probably through CaMKII and phosphorylation of RyR2, leading to an increase of the localized calcium releases (LCRs). These LCRs are then coupled to inward NCX current, and subsequently accelerate the DD and AP firing rate. At the same time, CaMKII may also increase the phosphorylation of PLN, which subsequently increases the activity of SERCA. The enhancement of the activity of both RyR2 and SERCA result in the acceleration of CaTs
    Figure Legend Snippet: Schematic diagram illustrating the mechanism through which TRPC7 positively regulates the automaticity of cardiomyocytes. G protein-coupled receptors (GPCRs) locating on the plasma membrane (PM) sense the external ligands such as hormones, neurotransmitters, and growth factors, transduce the signal to activate phospholipase C (PLC) which hydrolyzes the phosphatidylinositol 4,5-bisphosphate (PIP 2 ) into inositol trisphosphate (IP 3 ) and diacylglycerol (DAG). TRPC7 is then directly activated by DAG, mediating the Ca 2+ influx. Ca 2+ permeated through TRPC7 may activate the forward mode of the Na + -Ca 2+ exchanger (NCX), leading Na + influx and depolarization. This depolarization would then accelerate diastolic depolarization (DD) and increase AP firing rate. On the other hand, Ca 2+ permeated through TRPC7 may also increase the activity of ryanodine receptor 2 (RyR2) locating on the SR, probably through CaMKII and phosphorylation of RyR2, leading to an increase of the localized calcium releases (LCRs). These LCRs are then coupled to inward NCX current, and subsequently accelerate the DD and AP firing rate. At the same time, CaMKII may also increase the phosphorylation of PLN, which subsequently increases the activity of SERCA. The enhancement of the activity of both RyR2 and SERCA result in the acceleration of CaTs

    Techniques Used: Activity Assay

    ryr3  (Alomone Labs)


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    Alomone Labs ryr3
    Transcript levels for ryanodine receptor isoforms in mesenteric artery smooth muscle cells from young and old mice. Data are from 22 with permission.
    Ryr3, 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
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    Images

    1) Product Images from "Aging alters spontaneous and neurotransmitter-mediated Ca 2+ signaling in smooth muscle cells of mouse mesenteric arteries"

    Article Title: Aging alters spontaneous and neurotransmitter-mediated Ca 2+ signaling in smooth muscle cells of mouse mesenteric arteries

    Journal: Microcirculation (New York, N.Y. : 1994)

    doi: 10.1111/micc.12607

    Transcript levels for ryanodine receptor isoforms in mesenteric artery smooth muscle cells from young and old mice. Data are from 22 with permission.
    Figure Legend Snippet: Transcript levels for ryanodine receptor isoforms in mesenteric artery smooth muscle cells from young and old mice. Data are from 22 with permission.

    Techniques Used:

    Representative immunofluorescence images depicting green fluorescence for RyR1 (top rows), RyR2 (center rows) and RyR3 (bottom rows) in 3 separate SMCs from MAs of (A) young and (B) old mice, (C) SMCs incubated with respective blocking peptides, and (D) SMC with primary omitted. ToPro nuclear stain (blue) is included in all images. Scale bars = 20 μm and apply to all panels.
    Figure Legend Snippet: Representative immunofluorescence images depicting green fluorescence for RyR1 (top rows), RyR2 (center rows) and RyR3 (bottom rows) in 3 separate SMCs from MAs of (A) young and (B) old mice, (C) SMCs incubated with respective blocking peptides, and (D) SMC with primary omitted. ToPro nuclear stain (blue) is included in all images. Scale bars = 20 μm and apply to all panels.

    Techniques Used: Immunofluorescence, Fluorescence, Incubation, Blocking Assay, Staining

    ryr3  (Alomone Labs)


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    Alomone Labs ryr3
    Transcript levels for ryanodine receptor isoforms in mesenteric artery smooth muscle cells from young and old mice. Data are from 22 with permission.
    Ryr3, 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
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    Average 93 stars, based on 1 article reviews
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    ryr3 - by Bioz Stars, 2023-01
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    Images

    1) Product Images from "Aging alters spontaneous and neurotransmitter-mediated Ca 2+ signaling in smooth muscle cells of mouse mesenteric arteries"

    Article Title: Aging alters spontaneous and neurotransmitter-mediated Ca 2+ signaling in smooth muscle cells of mouse mesenteric arteries

    Journal: Microcirculation (New York, N.Y. : 1994)

    doi: 10.1111/micc.12607

    Transcript levels for ryanodine receptor isoforms in mesenteric artery smooth muscle cells from young and old mice. Data are from 22 with permission.
    Figure Legend Snippet: Transcript levels for ryanodine receptor isoforms in mesenteric artery smooth muscle cells from young and old mice. Data are from 22 with permission.

    Techniques Used:

    Representative immunofluorescence images depicting green fluorescence for RyR1 (top rows), RyR2 (center rows) and RyR3 (bottom rows) in 3 separate SMCs from MAs of (A) young and (B) old mice, (C) SMCs incubated with respective blocking peptides, and (D) SMC with primary omitted. ToPro nuclear stain (blue) is included in all images. Scale bars = 20 μm and apply to all panels.
    Figure Legend Snippet: Representative immunofluorescence images depicting green fluorescence for RyR1 (top rows), RyR2 (center rows) and RyR3 (bottom rows) in 3 separate SMCs from MAs of (A) young and (B) old mice, (C) SMCs incubated with respective blocking peptides, and (D) SMC with primary omitted. ToPro nuclear stain (blue) is included in all images. Scale bars = 20 μm and apply to all panels.

    Techniques Used: Immunofluorescence, Fluorescence, Incubation, Blocking Assay, Staining

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    Alomone Labs rabbit polyclonal ryr2
    CCN5 prevents cardiac fibrosis in mdx/utrn (±) mice. (A) Experimental scheme for panels (B–E) . mdx/utrn (±) mice were injected with AAV.9-Con or CCN5 into the tail vein, and hearts were harvested for experiments 8 weeks later. Age-matched WT mice are shown in comparison. (B) Hearts were sectioned and stained with trichrome. Blue areas indicate fibrotic tissue and red areas indicate normal tissue. (C) The ratio of fibrotic area over total tissue of the stained hearts was plotted. (D) Proteins obtained from cardiac tissue were immunoblotted with antibodies against CCN5, α-SMA, collagen I, SERCA2a, <t>RyR2,</t> NCX1, phosphorylated phospholamban (p-PLN), t-PLN and α-tubulin. (E) Protein bands on western blots were scanned and plotted. n = 6. * p < 0.05 and ** p < 0.01.
    Rabbit Polyclonal Ryr2, 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
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    Alomone Labs anti pser2808 ryr2
    Effects of macitentan on the calcium handling in LA in SHR. (a) Representative immunoblots showing expression of α1C, NCX1, CSQ and <t>RyR2</t> from SHR treated with MAC or DOX versus SHR-CTL (b-g) Quantification of immunoblots (N = 6–9 per group) (h) Representative immunoblots showing expression of SERCA2a, CaMKIIδ and PLB. Actin controls for pS16, pT17 and pS10 are not shown (i-m) Quantification of immunoblots (N = 6–9); # P < 0.05. Note that some blots/bands of housekeeping proteins used for normalization are identical (e.g. in (a) GAPDH blot/bands shown below CSQ and GAPDH blot/bands shown below RyR) as they are derived from the same membrane.
    Anti Pser2808 Ryr2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs anti ryanodine receptor 2 ryr2
    Discrete cAMP pools in adult mouse SAN cells (A) Diagrams highlighting localization and schematic representation of the Epac1-camps-based FRET biosensors (ICU3) in the cytosol (cyt; 1), plasma membrane (PM; 2), sarcoplasmic reticulum (SR; 3), myofilaments (MF; 4), and nucleus (nuc; 5). The ICU3 is linked to a Kras-derived sequence for PM localization, to a phospholamban (PLB)-derived sequence for SR localization, to a troponin T (TnT) for MF localization, and to a nuclear localization signal (NLS) sequence for nucleus localization. Exemplary super resolution images of adult wild-type mouse SAN cells expressing the indicated ICU3 biosensor in the cytosol (B), PM (C), SR (D), MF (E), and nucleus (F). The biosensor-associated fluorescence (YFP) is in magenta. Cells were immunostained with specific markers (in cyan) for the PM (caveolin 3), SR (ryanodine <t>receptor</t> <t>2</t> <t>[RyR2]),</t> MF (phalloidin; phal), and nucleus (DAPI). Merged images and corresponding line profile analysis (for dotted line) show high degree of overlap between the YFP fluorescence linked to the biosensor and the corresponding cellular marker in all cases, except in cells expressing the cytosolic sensor, as expected. Dotted squares highlight expanded regions in the solid squares. (G) Scatterplot of Pearson's correlation coefficient for cyt/cav3, PM/cav3, SR/RYR2, MF/phal, and nuc/DAPI (n > 8 SAN cells per condition). Kruskal-Wallis with Dunn's multiple comparisons test was used to test statistical differences in Pearson's correlation coefficient between non-target and targeted sensors. Scatterplot of the FRET ratio change in response to 10 μM forskolin (fsk) + 100 μM IBMX (H) and cAMP concentration-response curves (I) generated in HEK cells expressing the different ICU3 sensors (n > 5 cells per condition). For the cAMP concentration-response curves, cells expressing the different ICU3 sensors were exposed to increasing concentrations of the membrane-permeable cAMP analog 8CPT-cAMP. Kruskal-Wallis with Dunn's multiple comparisons test was used to compare fsk + IBMX responses, and the extra sum-of-squares F test was used to compare the cAMP EC 50 response between sensors. (J) Average FRET ratio traces (mean, solid lines; SEM, shadow) in response to 100 nM isoproterenol (iso) or 10 μM fsk from adult wild-type mouse SAN cells expressing the cytosolic, PM, SR, MF, or nuclear ICU3 biosensors (n > 5 cells from three preparations per condition). Scatterplots of ΔR/R 0 (K) and normalized (L) FRET responses after application of iso or fsk. Statistical differences were assessed with two-tailed Mann-Whitney test for comparisons between iso and fsk responses in (H) Statistical differences in fsk responses between the different biosensors in H were assessed with a Kruskal-Wallis with Dunn's multiple comparisons test. Statistical differences in normalized iso responses between the different groups were assessed using a one-way ANOVA with Tukey's multiple comparisons test. Significance (∗) was considered at P < 0.05. Exact p values are available in . Data represent mean ± SEM.
    Anti Ryanodine Receptor 2 Ryr2, 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
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    86
    Alomone Labs ryr2
    Ca2+ handling protein expression or phosphorylation at ZT4 and ZT14 in adult hearts. A-B. Phosphorylation of PLB at serine16 was essentially non-detectable following 5min perfusion (A) and 1hr perfusion (to match optical mapping data collection timepoint, B). Band labeled ‘Iso’ in (B) was a single heart perfused with isoproterenol for a positive control for pPLB. C-D. PLB phosphorylation was higher at the Thr17 site at ZT4 vs. ZT14 following 5min perfusion (C). However, phosphorylation levels were reduced and were not different between ZT4 and ZT14 following 1hr of perfusion (D). Band labeled ‘Iso’ in (D) was a single heart perfused with isoproterenol for a positive control for pPLB. E-G. Total PLB (E), SERCA2a (F), and <t>RyR2</t> (G) expression. α-tubulin was used as loading control. ZT4 n=4; ZT14 n=4; except RyR (G) where ZT4 n=3; ZT14 n=3. *p<0.05.
    Ryr2, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs antibody ryr1
    Flow cytometry detects <t>RYR1</t> in freshly isolated LMCs. Scatter plots of LMC suspensions plotted as side scatter (SSC-W) vs. fluorescent signals. Black horizontal lines denote the gate set. Fluorescent signal appearing above gate indicates positive staining. (A) Negative control (Neg) showing background FITC fluorescence of cell population with no antibody staining. Another LMC suspension was used to identify (B, left ) the LMC population stained by α-SMA-FITC ( red box ) and (B, right ) the gate set for non-specific staining of Alexa-Fluor 647 conjugated secondary antibody without primary antibodies within the α-SMA-FITC gated region. (C) Positive detection of RYR1 above the Alexa-Fluor 674 gate in LMCs isolated from rat mesenteric LVs. Data representative of five isolations each from male and female rats ( n = 10).
    Antibody Ryr1, 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
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    Alomone Labs ryr1 subtype
    Flow cytometry detects <t>RYR1</t> in freshly isolated LMCs. Scatter plots of LMC suspensions plotted as side scatter (SSC-W) vs. fluorescent signals. Black horizontal lines denote the gate set. Fluorescent signal appearing above gate indicates positive staining. (A) Negative control (Neg) showing background FITC fluorescence of cell population with no antibody staining. Another LMC suspension was used to identify (B, left ) the LMC population stained by α-SMA-FITC ( red box ) and (B, right ) the gate set for non-specific staining of Alexa-Fluor 647 conjugated secondary antibody without primary antibodies within the α-SMA-FITC gated region. (C) Positive detection of RYR1 above the Alexa-Fluor 674 gate in LMCs isolated from rat mesenteric LVs. Data representative of five isolations each from male and female rats ( n = 10).
    Ryr1 Subtype, 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
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    93
    Alomone Labs anti p s2814
    Knockdown of TRPC7 decreased the activity of <t>RyR2</t> and SERCA without affecting the activity of IP3R in mESC-CMs. Representative CaTs recorded from the control group a before and b after applying ryanodine. Representative CaTs recorded from TRPC7 knockdown group c before and d after applying ryanodine. e , f Bar charts showing the normalized frequency and the V max-upstroke calculated from a – d . The decrease of the frequency and the V max-upstroke caused by ryanodine was less in TRPC7 knockdown group. Representative CaTs recorded from the control group g before and h after applying thapsigargin (TG). Representative CaTs recorded from TRPC7 knockdown group i before and j after applying TG. k , l Bar charts showing the normalized frequency and V max-decay calculated from g – j . The decrease of the frequency and the V max-decay and amplitude caused by TG was less in TRPC7 knockdown group. Representative CaTs recorded from the control group m before and n after applying 2-aminoethoxydipheylborate (2-APB). Representative CaTs recorded from TRPC7 knockdown group o before and p after applying 2-APB. q , r Bar charts showing the normalized frequency and the V max-upstroke calculated from m – p . The decrease of the frequency and the V max-upstroke caused by 2-APB was similar between control and TRPC7 knockdown groups. Data were presented as mean ± SEM ( n = 10 cells; cells were from 3 independent batches of differentiation). * P < 0.05, ** P < 0.01 vs control
    Anti P S2814, 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
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    Alomone Labs ryr3
    Transcript levels for ryanodine receptor isoforms in mesenteric artery smooth muscle cells from young and old mice. Data are from 22 with permission.
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    Image Search Results


    CCN5 prevents cardiac fibrosis in mdx/utrn (±) mice. (A) Experimental scheme for panels (B–E) . mdx/utrn (±) mice were injected with AAV.9-Con or CCN5 into the tail vein, and hearts were harvested for experiments 8 weeks later. Age-matched WT mice are shown in comparison. (B) Hearts were sectioned and stained with trichrome. Blue areas indicate fibrotic tissue and red areas indicate normal tissue. (C) The ratio of fibrotic area over total tissue of the stained hearts was plotted. (D) Proteins obtained from cardiac tissue were immunoblotted with antibodies against CCN5, α-SMA, collagen I, SERCA2a, RyR2, NCX1, phosphorylated phospholamban (p-PLN), t-PLN and α-tubulin. (E) Protein bands on western blots were scanned and plotted. n = 6. * p < 0.05 and ** p < 0.01.

    Journal: Frontiers in Cardiovascular Medicine

    Article Title: Matricellular Protein CCN5 Gene Transfer Ameliorates Cardiac and Skeletal Dysfunction in mdx/utrn (±) Haploinsufficient Mice by Reducing Fibrosis and Upregulating Utrophin Expression

    doi: 10.3389/fcvm.2022.763544

    Figure Lengend Snippet: CCN5 prevents cardiac fibrosis in mdx/utrn (±) mice. (A) Experimental scheme for panels (B–E) . mdx/utrn (±) mice were injected with AAV.9-Con or CCN5 into the tail vein, and hearts were harvested for experiments 8 weeks later. Age-matched WT mice are shown in comparison. (B) Hearts were sectioned and stained with trichrome. Blue areas indicate fibrotic tissue and red areas indicate normal tissue. (C) The ratio of fibrotic area over total tissue of the stained hearts was plotted. (D) Proteins obtained from cardiac tissue were immunoblotted with antibodies against CCN5, α-SMA, collagen I, SERCA2a, RyR2, NCX1, phosphorylated phospholamban (p-PLN), t-PLN and α-tubulin. (E) Protein bands on western blots were scanned and plotted. n = 6. * p < 0.05 and ** p < 0.01.

    Article Snippet: Transferred blots were blocked with 5% non-fat skim milk and incubated with antibodies against mouse monoclonal CCN5 (1:1,000, Sigma, #WH0008839M9), mouse monoclonal α-SMA (1:1,000, Sigma-Aldrich, #A5228), goat polyclonal collagen I (1:1,000, Abcam, ab34710), rabbit polyclonal SERCA2a (1:1,000, a custom antibody from 21st Century Biochemicals), rabbit polyclonal RyR2 (1:1,000, Alomone Lab, ARR-002), rabbit monoclonal NCX1 (1:1,000, Abcam, EPR12739), rabbit polyclonal p-PLN (1:1,000, Badrilla, A010-12AP), mouse monoclonal t-PLN (1:1,000, Badrilla, A010-14), and mouse monoclonal α-tubulin (1:3,000, Santa Cruz, #sc-8035) for 12–16 h at 4°C.

    Techniques: Injection, Staining, Western Blot

    Effects of macitentan on the calcium handling in LA in SHR. (a) Representative immunoblots showing expression of α1C, NCX1, CSQ and RyR2 from SHR treated with MAC or DOX versus SHR-CTL (b-g) Quantification of immunoblots (N = 6–9 per group) (h) Representative immunoblots showing expression of SERCA2a, CaMKIIδ and PLB. Actin controls for pS16, pT17 and pS10 are not shown (i-m) Quantification of immunoblots (N = 6–9); # P < 0.05. Note that some blots/bands of housekeeping proteins used for normalization are identical (e.g. in (a) GAPDH blot/bands shown below CSQ and GAPDH blot/bands shown below RyR) as they are derived from the same membrane.

    Journal: International Journal of Cardiology. Heart & Vasculature

    Article Title: Anti-inflammatory effects of endothelin receptor blockade in left atrial tissue of spontaneously hypertensive rats

    doi: 10.1016/j.ijcha.2022.101088

    Figure Lengend Snippet: Effects of macitentan on the calcium handling in LA in SHR. (a) Representative immunoblots showing expression of α1C, NCX1, CSQ and RyR2 from SHR treated with MAC or DOX versus SHR-CTL (b-g) Quantification of immunoblots (N = 6–9 per group) (h) Representative immunoblots showing expression of SERCA2a, CaMKIIδ and PLB. Actin controls for pS16, pT17 and pS10 are not shown (i-m) Quantification of immunoblots (N = 6–9); # P < 0.05. Note that some blots/bands of housekeeping proteins used for normalization are identical (e.g. in (a) GAPDH blot/bands shown below CSQ and GAPDH blot/bands shown below RyR) as they are derived from the same membrane.

    Article Snippet: Quality of transfer was probed using Ponceau S. Primary antibodies used for detection of protein expression and phosphorylation were 1) on 4–20% gradient gels (BioRad, Germany): anti-CaMKII (50 kD, Badrilla, A010-55AP, 1:5,000), anti-CSQ (55 kD, Thermo Scientific, PA1-913, 1:2,500), anti-GAPDH (34 kD, Calbiochem, CB1001, 1:50,000), anti-IP 3 R2 (313 kD, Abcam, ab77838, 1:1,000), anti-NCX1 (120 kD, Thermo Scientific, MA1-4672, 1:1,000), anti-SERCA2a (100 kD, Badrilla, A010-20, 1:5,000), anti-RyR (565 kD, Thermo Scientific, MA3-916, 1:5,000), anti-pSer2808-RyR2 (Badrilla, A010-30, 1:5,000); 2) on 8% glycine gels: anti-Cav1.2a, i.e. cardiac type α1C subunit of the L-type Ca channel (250 kD, Alomone Labs, ACC-013, 1:200); and 3) on 14% tricine gels: anti-actin (44 kD, clone C4, MP Biomedicals #69100, 1:50,000), anti-PLB (25 kD, Badrilla, A010-14, 1:5,000), anti-Ser10-PLB (Badrilla, A010-10AP, 1:1,000), anti-pSer16-PLB (Badrilla, A010-12, 1:5,000), anti-pThr17-PLB (Badrilla, A010-13, 1:5,000).

    Techniques: Western Blot, Expressing, Derivative Assay

    Discrete cAMP pools in adult mouse SAN cells (A) Diagrams highlighting localization and schematic representation of the Epac1-camps-based FRET biosensors (ICU3) in the cytosol (cyt; 1), plasma membrane (PM; 2), sarcoplasmic reticulum (SR; 3), myofilaments (MF; 4), and nucleus (nuc; 5). The ICU3 is linked to a Kras-derived sequence for PM localization, to a phospholamban (PLB)-derived sequence for SR localization, to a troponin T (TnT) for MF localization, and to a nuclear localization signal (NLS) sequence for nucleus localization. Exemplary super resolution images of adult wild-type mouse SAN cells expressing the indicated ICU3 biosensor in the cytosol (B), PM (C), SR (D), MF (E), and nucleus (F). The biosensor-associated fluorescence (YFP) is in magenta. Cells were immunostained with specific markers (in cyan) for the PM (caveolin 3), SR (ryanodine receptor 2 [RyR2]), MF (phalloidin; phal), and nucleus (DAPI). Merged images and corresponding line profile analysis (for dotted line) show high degree of overlap between the YFP fluorescence linked to the biosensor and the corresponding cellular marker in all cases, except in cells expressing the cytosolic sensor, as expected. Dotted squares highlight expanded regions in the solid squares. (G) Scatterplot of Pearson's correlation coefficient for cyt/cav3, PM/cav3, SR/RYR2, MF/phal, and nuc/DAPI (n > 8 SAN cells per condition). Kruskal-Wallis with Dunn's multiple comparisons test was used to test statistical differences in Pearson's correlation coefficient between non-target and targeted sensors. Scatterplot of the FRET ratio change in response to 10 μM forskolin (fsk) + 100 μM IBMX (H) and cAMP concentration-response curves (I) generated in HEK cells expressing the different ICU3 sensors (n > 5 cells per condition). For the cAMP concentration-response curves, cells expressing the different ICU3 sensors were exposed to increasing concentrations of the membrane-permeable cAMP analog 8CPT-cAMP. Kruskal-Wallis with Dunn's multiple comparisons test was used to compare fsk + IBMX responses, and the extra sum-of-squares F test was used to compare the cAMP EC 50 response between sensors. (J) Average FRET ratio traces (mean, solid lines; SEM, shadow) in response to 100 nM isoproterenol (iso) or 10 μM fsk from adult wild-type mouse SAN cells expressing the cytosolic, PM, SR, MF, or nuclear ICU3 biosensors (n > 5 cells from three preparations per condition). Scatterplots of ΔR/R 0 (K) and normalized (L) FRET responses after application of iso or fsk. Statistical differences were assessed with two-tailed Mann-Whitney test for comparisons between iso and fsk responses in (H) Statistical differences in fsk responses between the different biosensors in H were assessed with a Kruskal-Wallis with Dunn's multiple comparisons test. Statistical differences in normalized iso responses between the different groups were assessed using a one-way ANOVA with Tukey's multiple comparisons test. Significance (∗) was considered at P < 0.05. Exact p values are available in . Data represent mean ± SEM.

    Journal: iScience

    Article Title: Deciphering cellular signals in adult mouse sinoatrial node cells

    doi: 10.1016/j.isci.2021.103693

    Figure Lengend Snippet: Discrete cAMP pools in adult mouse SAN cells (A) Diagrams highlighting localization and schematic representation of the Epac1-camps-based FRET biosensors (ICU3) in the cytosol (cyt; 1), plasma membrane (PM; 2), sarcoplasmic reticulum (SR; 3), myofilaments (MF; 4), and nucleus (nuc; 5). The ICU3 is linked to a Kras-derived sequence for PM localization, to a phospholamban (PLB)-derived sequence for SR localization, to a troponin T (TnT) for MF localization, and to a nuclear localization signal (NLS) sequence for nucleus localization. Exemplary super resolution images of adult wild-type mouse SAN cells expressing the indicated ICU3 biosensor in the cytosol (B), PM (C), SR (D), MF (E), and nucleus (F). The biosensor-associated fluorescence (YFP) is in magenta. Cells were immunostained with specific markers (in cyan) for the PM (caveolin 3), SR (ryanodine receptor 2 [RyR2]), MF (phalloidin; phal), and nucleus (DAPI). Merged images and corresponding line profile analysis (for dotted line) show high degree of overlap between the YFP fluorescence linked to the biosensor and the corresponding cellular marker in all cases, except in cells expressing the cytosolic sensor, as expected. Dotted squares highlight expanded regions in the solid squares. (G) Scatterplot of Pearson's correlation coefficient for cyt/cav3, PM/cav3, SR/RYR2, MF/phal, and nuc/DAPI (n > 8 SAN cells per condition). Kruskal-Wallis with Dunn's multiple comparisons test was used to test statistical differences in Pearson's correlation coefficient between non-target and targeted sensors. Scatterplot of the FRET ratio change in response to 10 μM forskolin (fsk) + 100 μM IBMX (H) and cAMP concentration-response curves (I) generated in HEK cells expressing the different ICU3 sensors (n > 5 cells per condition). For the cAMP concentration-response curves, cells expressing the different ICU3 sensors were exposed to increasing concentrations of the membrane-permeable cAMP analog 8CPT-cAMP. Kruskal-Wallis with Dunn's multiple comparisons test was used to compare fsk + IBMX responses, and the extra sum-of-squares F test was used to compare the cAMP EC 50 response between sensors. (J) Average FRET ratio traces (mean, solid lines; SEM, shadow) in response to 100 nM isoproterenol (iso) or 10 μM fsk from adult wild-type mouse SAN cells expressing the cytosolic, PM, SR, MF, or nuclear ICU3 biosensors (n > 5 cells from three preparations per condition). Scatterplots of ΔR/R 0 (K) and normalized (L) FRET responses after application of iso or fsk. Statistical differences were assessed with two-tailed Mann-Whitney test for comparisons between iso and fsk responses in (H) Statistical differences in fsk responses between the different biosensors in H were assessed with a Kruskal-Wallis with Dunn's multiple comparisons test. Statistical differences in normalized iso responses between the different groups were assessed using a one-way ANOVA with Tukey's multiple comparisons test. Significance (∗) was considered at P < 0.05. Exact p values are available in . Data represent mean ± SEM.

    Article Snippet: For experiments in , cells infected with the different ICU3 biosensors were incubated with an anti-caveolin 3 antibody (1:200, Thermo-Fisher Scientific PA1-066) to label the PM, an anti-ryanodine receptor 2 (RyR2) antibody (1:200, Alomone Labs ARR-002) to label the SR, an Alexa Fluor 647 Phalloidin (1.3 U; Thermo-Fisher Scientific A22287) to label MF, or DAPI from the ProLong Diamond Antifade Mountant (Thermo-Fisher Scientific) to label the nucleus.

    Techniques: Derivative Assay, Sequencing, Expressing, Fluorescence, Marker, Concentration Assay, Generated, Two Tailed Test, MANN-WHITNEY

    Journal: iScience

    Article Title: Deciphering cellular signals in adult mouse sinoatrial node cells

    doi: 10.1016/j.isci.2021.103693

    Figure Lengend Snippet:

    Article Snippet: For experiments in , cells infected with the different ICU3 biosensors were incubated with an anti-caveolin 3 antibody (1:200, Thermo-Fisher Scientific PA1-066) to label the PM, an anti-ryanodine receptor 2 (RyR2) antibody (1:200, Alomone Labs ARR-002) to label the SR, an Alexa Fluor 647 Phalloidin (1.3 U; Thermo-Fisher Scientific A22287) to label MF, or DAPI from the ProLong Diamond Antifade Mountant (Thermo-Fisher Scientific) to label the nucleus.

    Techniques: Recombinant, Software

    Ca2+ handling protein expression or phosphorylation at ZT4 and ZT14 in adult hearts. A-B. Phosphorylation of PLB at serine16 was essentially non-detectable following 5min perfusion (A) and 1hr perfusion (to match optical mapping data collection timepoint, B). Band labeled ‘Iso’ in (B) was a single heart perfused with isoproterenol for a positive control for pPLB. C-D. PLB phosphorylation was higher at the Thr17 site at ZT4 vs. ZT14 following 5min perfusion (C). However, phosphorylation levels were reduced and were not different between ZT4 and ZT14 following 1hr of perfusion (D). Band labeled ‘Iso’ in (D) was a single heart perfused with isoproterenol for a positive control for pPLB. E-G. Total PLB (E), SERCA2a (F), and RyR2 (G) expression. α-tubulin was used as loading control. ZT4 n=4; ZT14 n=4; except RyR (G) where ZT4 n=3; ZT14 n=3. *p<0.05.

    Journal: Circulation. Arrhythmia and electrophysiology

    Article Title: Aging Disrupts Normal Time-of-day Variation in Cardiac Electrophysiology

    doi: 10.1161/CIRCEP.119.008093

    Figure Lengend Snippet: Ca2+ handling protein expression or phosphorylation at ZT4 and ZT14 in adult hearts. A-B. Phosphorylation of PLB at serine16 was essentially non-detectable following 5min perfusion (A) and 1hr perfusion (to match optical mapping data collection timepoint, B). Band labeled ‘Iso’ in (B) was a single heart perfused with isoproterenol for a positive control for pPLB. C-D. PLB phosphorylation was higher at the Thr17 site at ZT4 vs. ZT14 following 5min perfusion (C). However, phosphorylation levels were reduced and were not different between ZT4 and ZT14 following 1hr of perfusion (D). Band labeled ‘Iso’ in (D) was a single heart perfused with isoproterenol for a positive control for pPLB. E-G. Total PLB (E), SERCA2a (F), and RyR2 (G) expression. α-tubulin was used as loading control. ZT4 n=4; ZT14 n=4; except RyR (G) where ZT4 n=3; ZT14 n=3. *p<0.05.

    Article Snippet: 25 Anti-SERCA (1:5000, A010–20, Badrilla), RyR2 (1:2500, ARR002, Alomone), PLB (1:5000, A010–14, Badrilla), PLB-Thr17 (1:5000, A010–13, Badrilla), PLB-Ser16 (1:5000, A010–12, Badrilla), and α-tubulin (1:1000, ab52866, Abcam) were used as primary antibodies.

    Techniques: Expressing, Labeling, Positive Control

    Flow cytometry detects RYR1 in freshly isolated LMCs. Scatter plots of LMC suspensions plotted as side scatter (SSC-W) vs. fluorescent signals. Black horizontal lines denote the gate set. Fluorescent signal appearing above gate indicates positive staining. (A) Negative control (Neg) showing background FITC fluorescence of cell population with no antibody staining. Another LMC suspension was used to identify (B, left ) the LMC population stained by α-SMA-FITC ( red box ) and (B, right ) the gate set for non-specific staining of Alexa-Fluor 647 conjugated secondary antibody without primary antibodies within the α-SMA-FITC gated region. (C) Positive detection of RYR1 above the Alexa-Fluor 674 gate in LMCs isolated from rat mesenteric LVs. Data representative of five isolations each from male and female rats ( n = 10).

    Journal: Frontiers in Pharmacology

    Article Title: Dantrolene Prevents the Lymphostasis Caused by Doxorubicin in the Rat Mesenteric Circulation

    doi: 10.3389/fphar.2021.727526

    Figure Lengend Snippet: Flow cytometry detects RYR1 in freshly isolated LMCs. Scatter plots of LMC suspensions plotted as side scatter (SSC-W) vs. fluorescent signals. Black horizontal lines denote the gate set. Fluorescent signal appearing above gate indicates positive staining. (A) Negative control (Neg) showing background FITC fluorescence of cell population with no antibody staining. Another LMC suspension was used to identify (B, left ) the LMC population stained by α-SMA-FITC ( red box ) and (B, right ) the gate set for non-specific staining of Alexa-Fluor 647 conjugated secondary antibody without primary antibodies within the α-SMA-FITC gated region. (C) Positive detection of RYR1 above the Alexa-Fluor 674 gate in LMCs isolated from rat mesenteric LVs. Data representative of five isolations each from male and female rats ( n = 10).

    Article Snippet: The membrane was first incubated in tris-buffered saline (TBS) containing 0.05% Tween-20 and 5% non-fat dry milk to reduce non-specific antibody binding, then incubated overnight at 4°C with primary antibody RYR1 (1:1,000, Alomone Labs, Jerusalem, Israel) prepared in TBS containing 0.05% Tween-20 and 5% non-fat dry milk.

    Techniques: Flow Cytometry, Isolation, Staining, Negative Control, Fluorescence

    RYR1 antibody selectivity. (A) Western blot shows positive detection of RYR1 (∼565 kD) in rat skeletal muscle lysate and no detection in rat heart or brain lysate. (B,C) Contour plots of RYR1 antibody testing. (B) Red population represents positive detection of RYR1 subtype in male LMCs. Grey population represents the LMC suspension containing no primary antibody. (C) No detection in male LMCs after co-incubation with competing peptide (CP; 1:50 dilution) for the RYR1 antibody. Data representative of three isolations for each sex ( n = 6).

    Journal: Frontiers in Pharmacology

    Article Title: Dantrolene Prevents the Lymphostasis Caused by Doxorubicin in the Rat Mesenteric Circulation

    doi: 10.3389/fphar.2021.727526

    Figure Lengend Snippet: RYR1 antibody selectivity. (A) Western blot shows positive detection of RYR1 (∼565 kD) in rat skeletal muscle lysate and no detection in rat heart or brain lysate. (B,C) Contour plots of RYR1 antibody testing. (B) Red population represents positive detection of RYR1 subtype in male LMCs. Grey population represents the LMC suspension containing no primary antibody. (C) No detection in male LMCs after co-incubation with competing peptide (CP; 1:50 dilution) for the RYR1 antibody. Data representative of three isolations for each sex ( n = 6).

    Article Snippet: The membrane was first incubated in tris-buffered saline (TBS) containing 0.05% Tween-20 and 5% non-fat dry milk to reduce non-specific antibody binding, then incubated overnight at 4°C with primary antibody RYR1 (1:1,000, Alomone Labs, Jerusalem, Israel) prepared in TBS containing 0.05% Tween-20 and 5% non-fat dry milk.

    Techniques: Western Blot, Incubation

    Flow cytometry detects RYR1 in freshly isolated LMCs. Scatter plots of LMC suspensions plotted as side scatter (SSC-W) vs. fluorescent signals. Black horizontal lines denote the gate set. Fluorescent signal appearing above gate indicates positive staining. (A) Negative control (Neg) showing background FITC fluorescence of cell population with no antibody staining. Another LMC suspension was used to identify (B, left ) the LMC population stained by α-SMA-FITC ( red box ) and (B, right ) the gate set for non-specific staining of Alexa-Fluor 647 conjugated secondary antibody without primary antibodies within the α-SMA-FITC gated region. (C) Positive detection of RYR1 above the Alexa-Fluor 674 gate in LMCs isolated from rat mesenteric LVs. Data representative of five isolations each from male and female rats ( n = 10).

    Journal: Frontiers in Pharmacology

    Article Title: Dantrolene Prevents the Lymphostasis Caused by Doxorubicin in the Rat Mesenteric Circulation

    doi: 10.3389/fphar.2021.727526

    Figure Lengend Snippet: Flow cytometry detects RYR1 in freshly isolated LMCs. Scatter plots of LMC suspensions plotted as side scatter (SSC-W) vs. fluorescent signals. Black horizontal lines denote the gate set. Fluorescent signal appearing above gate indicates positive staining. (A) Negative control (Neg) showing background FITC fluorescence of cell population with no antibody staining. Another LMC suspension was used to identify (B, left ) the LMC population stained by α-SMA-FITC ( red box ) and (B, right ) the gate set for non-specific staining of Alexa-Fluor 647 conjugated secondary antibody without primary antibodies within the α-SMA-FITC gated region. (C) Positive detection of RYR1 above the Alexa-Fluor 674 gate in LMCs isolated from rat mesenteric LVs. Data representative of five isolations each from male and female rats ( n = 10).

    Article Snippet: Then cells were divided into groups for incubation with and without primary antibody targeting the RYR1 subtype (Alomone Labs, Jerusalem, Israel; ARR-001) at a dilution of 1:200 and rotated at 4°C overnight.

    Techniques: Flow Cytometry, Isolation, Staining, Negative Control, Fluorescence

    RYR1 antibody selectivity. (A) Western blot shows positive detection of RYR1 (∼565 kD) in rat skeletal muscle lysate and no detection in rat heart or brain lysate. (B,C) Contour plots of RYR1 antibody testing. (B) Red population represents positive detection of RYR1 subtype in male LMCs. Grey population represents the LMC suspension containing no primary antibody. (C) No detection in male LMCs after co-incubation with competing peptide (CP; 1:50 dilution) for the RYR1 antibody. Data representative of three isolations for each sex ( n = 6).

    Journal: Frontiers in Pharmacology

    Article Title: Dantrolene Prevents the Lymphostasis Caused by Doxorubicin in the Rat Mesenteric Circulation

    doi: 10.3389/fphar.2021.727526

    Figure Lengend Snippet: RYR1 antibody selectivity. (A) Western blot shows positive detection of RYR1 (∼565 kD) in rat skeletal muscle lysate and no detection in rat heart or brain lysate. (B,C) Contour plots of RYR1 antibody testing. (B) Red population represents positive detection of RYR1 subtype in male LMCs. Grey population represents the LMC suspension containing no primary antibody. (C) No detection in male LMCs after co-incubation with competing peptide (CP; 1:50 dilution) for the RYR1 antibody. Data representative of three isolations for each sex ( n = 6).

    Article Snippet: Then cells were divided into groups for incubation with and without primary antibody targeting the RYR1 subtype (Alomone Labs, Jerusalem, Israel; ARR-001) at a dilution of 1:200 and rotated at 4°C overnight.

    Techniques: Western Blot, Incubation

    Knockdown of TRPC7 decreased the activity of RyR2 and SERCA without affecting the activity of IP3R in mESC-CMs. Representative CaTs recorded from the control group a before and b after applying ryanodine. Representative CaTs recorded from TRPC7 knockdown group c before and d after applying ryanodine. e , f Bar charts showing the normalized frequency and the V max-upstroke calculated from a – d . The decrease of the frequency and the V max-upstroke caused by ryanodine was less in TRPC7 knockdown group. Representative CaTs recorded from the control group g before and h after applying thapsigargin (TG). Representative CaTs recorded from TRPC7 knockdown group i before and j after applying TG. k , l Bar charts showing the normalized frequency and V max-decay calculated from g – j . The decrease of the frequency and the V max-decay and amplitude caused by TG was less in TRPC7 knockdown group. Representative CaTs recorded from the control group m before and n after applying 2-aminoethoxydipheylborate (2-APB). Representative CaTs recorded from TRPC7 knockdown group o before and p after applying 2-APB. q , r Bar charts showing the normalized frequency and the V max-upstroke calculated from m – p . The decrease of the frequency and the V max-upstroke caused by 2-APB was similar between control and TRPC7 knockdown groups. Data were presented as mean ± SEM ( n = 10 cells; cells were from 3 independent batches of differentiation). * P < 0.05, ** P < 0.01 vs control

    Journal: Stem Cell Research & Therapy

    Article Title: TRPC7 regulates the electrophysiological functions of embryonic stem cell-derived cardiomyocytes

    doi: 10.1186/s13287-021-02308-7

    Figure Lengend Snippet: Knockdown of TRPC7 decreased the activity of RyR2 and SERCA without affecting the activity of IP3R in mESC-CMs. Representative CaTs recorded from the control group a before and b after applying ryanodine. Representative CaTs recorded from TRPC7 knockdown group c before and d after applying ryanodine. e , f Bar charts showing the normalized frequency and the V max-upstroke calculated from a – d . The decrease of the frequency and the V max-upstroke caused by ryanodine was less in TRPC7 knockdown group. Representative CaTs recorded from the control group g before and h after applying thapsigargin (TG). Representative CaTs recorded from TRPC7 knockdown group i before and j after applying TG. k , l Bar charts showing the normalized frequency and V max-decay calculated from g – j . The decrease of the frequency and the V max-decay and amplitude caused by TG was less in TRPC7 knockdown group. Representative CaTs recorded from the control group m before and n after applying 2-aminoethoxydipheylborate (2-APB). Representative CaTs recorded from TRPC7 knockdown group o before and p after applying 2-APB. q , r Bar charts showing the normalized frequency and the V max-upstroke calculated from m – p . The decrease of the frequency and the V max-upstroke caused by 2-APB was similar between control and TRPC7 knockdown groups. Data were presented as mean ± SEM ( n = 10 cells; cells were from 3 independent batches of differentiation). * P < 0.05, ** P < 0.01 vs control

    Article Snippet: Antibodies used were anti-TRPC7 1:500 (HPA031126, Sigma), anti-β-tubulin 1:1000 (15,115, Cell Signaling, Danvers, Massachusetts, USA), anti-HCN4 1:200 (APC-052, Alomone), anti-Cav1.3 1:100 (ACC-005, Alomone), anti-Cav3.1 1:200 (ACC-021, Alomone), anti-Cav3.2 1:200 (ACC-025, Alomone), anti-RyR2 1:1000 (MA3-916, Invitrogen), anti-SERCA 1:200 (sc-30110, Santa Cruz), anti-IP3R 1:500 (ACC-019, Alomone), anti-p(S2814)RyR2 1:5000 (A010-31, Badrilla), anti-phospholamban (PLN) 1:1000 (A010-14, Badrilla), anti-p(T17) PLN 1:5000 (A010-13, Badrilla), HRP-conjugated goat anti-rabbit secondary antibody 1:5000 (Dako, Zug, Switzerland), and HRP-conjugated goat anti-mouse secondary antibody 1:5000 (Dako).

    Techniques: Activity Assay

    Knockdown of TRPC7 decreased while overexpression of TRPC7 increased the phosphorylation of RyR2 and PLN, respectively, in NRVMs. Western blotting showing the expression of a total RyR2, b p(S2814)RyR2, c total PLN, and d p(T17) PLN in NRVMs infected with different adenoviruses to knockdown or overexpress TRPC7. e – h Bar charts showing the quantification of each protein from a – d . To eliminate the loading bias, intensity of each target protein was normalized to that of its corresponding β-tubulin. The change of TRPC7 expression did not alter the expression of total RyR2 and PLN. However, knockdown of TRPC7 decreased while overexpression of TRPC7 increased the phosphorylation form of RyR2 [p(S2814)RyR2] and PLN [p(T17)PLN]. Data were presented as mean ± SEM ( n = 4). * P < 0.05, *** P < 0.001 vs corresponding controls

    Journal: Stem Cell Research & Therapy

    Article Title: TRPC7 regulates the electrophysiological functions of embryonic stem cell-derived cardiomyocytes

    doi: 10.1186/s13287-021-02308-7

    Figure Lengend Snippet: Knockdown of TRPC7 decreased while overexpression of TRPC7 increased the phosphorylation of RyR2 and PLN, respectively, in NRVMs. Western blotting showing the expression of a total RyR2, b p(S2814)RyR2, c total PLN, and d p(T17) PLN in NRVMs infected with different adenoviruses to knockdown or overexpress TRPC7. e – h Bar charts showing the quantification of each protein from a – d . To eliminate the loading bias, intensity of each target protein was normalized to that of its corresponding β-tubulin. The change of TRPC7 expression did not alter the expression of total RyR2 and PLN. However, knockdown of TRPC7 decreased while overexpression of TRPC7 increased the phosphorylation form of RyR2 [p(S2814)RyR2] and PLN [p(T17)PLN]. Data were presented as mean ± SEM ( n = 4). * P < 0.05, *** P < 0.001 vs corresponding controls

    Article Snippet: Antibodies used were anti-TRPC7 1:500 (HPA031126, Sigma), anti-β-tubulin 1:1000 (15,115, Cell Signaling, Danvers, Massachusetts, USA), anti-HCN4 1:200 (APC-052, Alomone), anti-Cav1.3 1:100 (ACC-005, Alomone), anti-Cav3.1 1:200 (ACC-021, Alomone), anti-Cav3.2 1:200 (ACC-025, Alomone), anti-RyR2 1:1000 (MA3-916, Invitrogen), anti-SERCA 1:200 (sc-30110, Santa Cruz), anti-IP3R 1:500 (ACC-019, Alomone), anti-p(S2814)RyR2 1:5000 (A010-31, Badrilla), anti-phospholamban (PLN) 1:1000 (A010-14, Badrilla), anti-p(T17) PLN 1:5000 (A010-13, Badrilla), HRP-conjugated goat anti-rabbit secondary antibody 1:5000 (Dako, Zug, Switzerland), and HRP-conjugated goat anti-mouse secondary antibody 1:5000 (Dako).

    Techniques: Over Expression, Western Blot, Expressing, Infection

    Schematic diagram illustrating the mechanism through which TRPC7 positively regulates the automaticity of cardiomyocytes. G protein-coupled receptors (GPCRs) locating on the plasma membrane (PM) sense the external ligands such as hormones, neurotransmitters, and growth factors, transduce the signal to activate phospholipase C (PLC) which hydrolyzes the phosphatidylinositol 4,5-bisphosphate (PIP 2 ) into inositol trisphosphate (IP 3 ) and diacylglycerol (DAG). TRPC7 is then directly activated by DAG, mediating the Ca 2+ influx. Ca 2+ permeated through TRPC7 may activate the forward mode of the Na + -Ca 2+ exchanger (NCX), leading Na + influx and depolarization. This depolarization would then accelerate diastolic depolarization (DD) and increase AP firing rate. On the other hand, Ca 2+ permeated through TRPC7 may also increase the activity of ryanodine receptor 2 (RyR2) locating on the SR, probably through CaMKII and phosphorylation of RyR2, leading to an increase of the localized calcium releases (LCRs). These LCRs are then coupled to inward NCX current, and subsequently accelerate the DD and AP firing rate. At the same time, CaMKII may also increase the phosphorylation of PLN, which subsequently increases the activity of SERCA. The enhancement of the activity of both RyR2 and SERCA result in the acceleration of CaTs

    Journal: Stem Cell Research & Therapy

    Article Title: TRPC7 regulates the electrophysiological functions of embryonic stem cell-derived cardiomyocytes

    doi: 10.1186/s13287-021-02308-7

    Figure Lengend Snippet: Schematic diagram illustrating the mechanism through which TRPC7 positively regulates the automaticity of cardiomyocytes. G protein-coupled receptors (GPCRs) locating on the plasma membrane (PM) sense the external ligands such as hormones, neurotransmitters, and growth factors, transduce the signal to activate phospholipase C (PLC) which hydrolyzes the phosphatidylinositol 4,5-bisphosphate (PIP 2 ) into inositol trisphosphate (IP 3 ) and diacylglycerol (DAG). TRPC7 is then directly activated by DAG, mediating the Ca 2+ influx. Ca 2+ permeated through TRPC7 may activate the forward mode of the Na + -Ca 2+ exchanger (NCX), leading Na + influx and depolarization. This depolarization would then accelerate diastolic depolarization (DD) and increase AP firing rate. On the other hand, Ca 2+ permeated through TRPC7 may also increase the activity of ryanodine receptor 2 (RyR2) locating on the SR, probably through CaMKII and phosphorylation of RyR2, leading to an increase of the localized calcium releases (LCRs). These LCRs are then coupled to inward NCX current, and subsequently accelerate the DD and AP firing rate. At the same time, CaMKII may also increase the phosphorylation of PLN, which subsequently increases the activity of SERCA. The enhancement of the activity of both RyR2 and SERCA result in the acceleration of CaTs

    Article Snippet: Antibodies used were anti-TRPC7 1:500 (HPA031126, Sigma), anti-β-tubulin 1:1000 (15,115, Cell Signaling, Danvers, Massachusetts, USA), anti-HCN4 1:200 (APC-052, Alomone), anti-Cav1.3 1:100 (ACC-005, Alomone), anti-Cav3.1 1:200 (ACC-021, Alomone), anti-Cav3.2 1:200 (ACC-025, Alomone), anti-RyR2 1:1000 (MA3-916, Invitrogen), anti-SERCA 1:200 (sc-30110, Santa Cruz), anti-IP3R 1:500 (ACC-019, Alomone), anti-p(S2814)RyR2 1:5000 (A010-31, Badrilla), anti-phospholamban (PLN) 1:1000 (A010-14, Badrilla), anti-p(T17) PLN 1:5000 (A010-13, Badrilla), HRP-conjugated goat anti-rabbit secondary antibody 1:5000 (Dako, Zug, Switzerland), and HRP-conjugated goat anti-mouse secondary antibody 1:5000 (Dako).

    Techniques: Activity Assay

    Transcript levels for ryanodine receptor isoforms in mesenteric artery smooth muscle cells from young and old mice. Data are from 22 with permission.

    Journal: Microcirculation (New York, N.Y. : 1994)

    Article Title: Aging alters spontaneous and neurotransmitter-mediated Ca 2+ signaling in smooth muscle cells of mouse mesenteric arteries

    doi: 10.1111/micc.12607

    Figure Lengend Snippet: Transcript levels for ryanodine receptor isoforms in mesenteric artery smooth muscle cells from young and old mice. Data are from 22 with permission.

    Article Snippet: Thus prepared, slides were incubated 60 min in primary antibody [Alamone Labs, 1:250] for RyR1 (Cat. #ARR-001), RyR2 (Cat. #ARR-002), or RyR3 (Cat. #ARR-003).

    Techniques:

    Representative immunofluorescence images depicting green fluorescence for RyR1 (top rows), RyR2 (center rows) and RyR3 (bottom rows) in 3 separate SMCs from MAs of (A) young and (B) old mice, (C) SMCs incubated with respective blocking peptides, and (D) SMC with primary omitted. ToPro nuclear stain (blue) is included in all images. Scale bars = 20 μm and apply to all panels.

    Journal: Microcirculation (New York, N.Y. : 1994)

    Article Title: Aging alters spontaneous and neurotransmitter-mediated Ca 2+ signaling in smooth muscle cells of mouse mesenteric arteries

    doi: 10.1111/micc.12607

    Figure Lengend Snippet: Representative immunofluorescence images depicting green fluorescence for RyR1 (top rows), RyR2 (center rows) and RyR3 (bottom rows) in 3 separate SMCs from MAs of (A) young and (B) old mice, (C) SMCs incubated with respective blocking peptides, and (D) SMC with primary omitted. ToPro nuclear stain (blue) is included in all images. Scale bars = 20 μm and apply to all panels.

    Article Snippet: Thus prepared, slides were incubated 60 min in primary antibody [Alamone Labs, 1:250] for RyR1 (Cat. #ARR-001), RyR2 (Cat. #ARR-002), or RyR3 (Cat. #ARR-003).

    Techniques: Immunofluorescence, Fluorescence, Incubation, Blocking Assay, Staining