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btp 2  (MedChemExpress)
93
MedChemExpress btp 2
Effects of Drebrin on the different stages of PRV infection. ( A ) Schematic timeline for the time-of-addition assays. ( B <t>)</t> <t>BTP−2</t> (600 nM) was co-incubated with PRV-QXX (MOI = 10) at 4 °C for 2 h to simulate virus adsorption under low-temperature conditions in PK−15 cells. Unbound viral particles were removed, viral genomic DNA was extracted, and the copy numbers of the viral genome were evaluated by qRT-PCR. ( C ) PK−15 cells were firstly incubated with PRV-QXX (MOI = 10) at 4 °C for 1 h to allow virus adsorption. Subsequently, unbound virus was removed, and medium containing 600 nM BTP−2 was added. Cells were treated at 37 °C for 1 h, and viral genome copy number was evaluated by qRT-PCR. ( D ) PK−15 cells were incubated with PRV−QXX (MOI = 1) at 4 °C for 1 h, and the inoculum was then removed and replaced with maintenance medium containing 2% FBS/DMEM. Cells were incubated at 37 °C for 12 h to permit viral entry and early replication events. Subsequently, BTP2 was added, and cells were incubated at 37 °C for another 12 h. Then, the virus was collected by being frozen and thawed three times, and the TCID 50 assay was performed to evaluate the virus titer. ( E ) PK−15 cells were treated as in ( D ): the supernatant was collected and TCID 50 was performed to evaluate the extracellular viral yield. ( F ) The sgCtrl and sgDBN cells were incubated with PRV-QXX at 4 °C for 2 h, cells were rinsed 3 times with ice-cold PBS and incubated with maintenance solution at 37 °C for 30 min, and then the PRV genome copy number was determined by RT-qPCR. ( G ) Cells were treated as in ( D ): viruses were harvested with three freeze–thaw cycles, and then the viral titers were analyzed by TCID 50 assay. ( H , I ) The sgCtrl and sgDBN cells were incubated with PRV−QXX at 37 °C for 1 h, cells were rinsed three times with PBS, and the medium was replaced with 2% FBS/DMEM to maintain growth for 12 h and 24 h. Then, the cells and supernatant were collected to analyze the intracellular and extracellular viral yield by the TCID 50 assay, respectively. Significance levels relative to the corresponding control were denoted as non-significant * p < 0.05, ** p < 0.01, and *** p < 0.001.
Btp 2, supplied by MedChemExpress, 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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2-btp  (Markstein Sichtec Medical)
90
Markstein Sichtec Medical 2-btp
Effects of Drebrin on the different stages of PRV infection. ( A ) Schematic timeline for the time-of-addition assays. ( B <t>)</t> <t>BTP−2</t> (600 nM) was co-incubated with PRV-QXX (MOI = 10) at 4 °C for 2 h to simulate virus adsorption under low-temperature conditions in PK−15 cells. Unbound viral particles were removed, viral genomic DNA was extracted, and the copy numbers of the viral genome were evaluated by qRT-PCR. ( C ) PK−15 cells were firstly incubated with PRV-QXX (MOI = 10) at 4 °C for 1 h to allow virus adsorption. Subsequently, unbound virus was removed, and medium containing 600 nM BTP−2 was added. Cells were treated at 37 °C for 1 h, and viral genome copy number was evaluated by qRT-PCR. ( D ) PK−15 cells were incubated with PRV−QXX (MOI = 1) at 4 °C for 1 h, and the inoculum was then removed and replaced with maintenance medium containing 2% FBS/DMEM. Cells were incubated at 37 °C for 12 h to permit viral entry and early replication events. Subsequently, BTP2 was added, and cells were incubated at 37 °C for another 12 h. Then, the virus was collected by being frozen and thawed three times, and the TCID 50 assay was performed to evaluate the virus titer. ( E ) PK−15 cells were treated as in ( D ): the supernatant was collected and TCID 50 was performed to evaluate the extracellular viral yield. ( F ) The sgCtrl and sgDBN cells were incubated with PRV-QXX at 4 °C for 2 h, cells were rinsed 3 times with ice-cold PBS and incubated with maintenance solution at 37 °C for 30 min, and then the PRV genome copy number was determined by RT-qPCR. ( G ) Cells were treated as in ( D ): viruses were harvested with three freeze–thaw cycles, and then the viral titers were analyzed by TCID 50 assay. ( H , I ) The sgCtrl and sgDBN cells were incubated with PRV−QXX at 37 °C for 1 h, cells were rinsed three times with PBS, and the medium was replaced with 2% FBS/DMEM to maintain growth for 12 h and 24 h. Then, the cells and supernatant were collected to analyze the intracellular and extracellular viral yield by the TCID 50 assay, respectively. Significance levels relative to the corresponding control were denoted as non-significant * p < 0.05, ** p < 0.01, and *** p < 0.001.
2 Btp, supplied by Markstein Sichtec Medical, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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btp-2  (Tocris)
90
Tocris btp-2
Effects of Drebrin on the different stages of PRV infection. ( A ) Schematic timeline for the time-of-addition assays. ( B <t>)</t> <t>BTP−2</t> (600 nM) was co-incubated with PRV-QXX (MOI = 10) at 4 °C for 2 h to simulate virus adsorption under low-temperature conditions in PK−15 cells. Unbound viral particles were removed, viral genomic DNA was extracted, and the copy numbers of the viral genome were evaluated by qRT-PCR. ( C ) PK−15 cells were firstly incubated with PRV-QXX (MOI = 10) at 4 °C for 1 h to allow virus adsorption. Subsequently, unbound virus was removed, and medium containing 600 nM BTP−2 was added. Cells were treated at 37 °C for 1 h, and viral genome copy number was evaluated by qRT-PCR. ( D ) PK−15 cells were incubated with PRV−QXX (MOI = 1) at 4 °C for 1 h, and the inoculum was then removed and replaced with maintenance medium containing 2% FBS/DMEM. Cells were incubated at 37 °C for 12 h to permit viral entry and early replication events. Subsequently, BTP2 was added, and cells were incubated at 37 °C for another 12 h. Then, the virus was collected by being frozen and thawed three times, and the TCID 50 assay was performed to evaluate the virus titer. ( E ) PK−15 cells were treated as in ( D ): the supernatant was collected and TCID 50 was performed to evaluate the extracellular viral yield. ( F ) The sgCtrl and sgDBN cells were incubated with PRV-QXX at 4 °C for 2 h, cells were rinsed 3 times with ice-cold PBS and incubated with maintenance solution at 37 °C for 30 min, and then the PRV genome copy number was determined by RT-qPCR. ( G ) Cells were treated as in ( D ): viruses were harvested with three freeze–thaw cycles, and then the viral titers were analyzed by TCID 50 assay. ( H , I ) The sgCtrl and sgDBN cells were incubated with PRV−QXX at 37 °C for 1 h, cells were rinsed three times with PBS, and the medium was replaced with 2% FBS/DMEM to maintain growth for 12 h and 24 h. Then, the cells and supernatant were collected to analyze the intracellular and extracellular viral yield by the TCID 50 assay, respectively. Significance levels relative to the corresponding control were denoted as non-significant * p < 0.05, ** p < 0.01, and *** p < 0.001.
Btp 2, supplied by Tocris, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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ym-58483 known btp-2  (Millipore)
90
Millipore ym-58483 known btp-2
Effects of Drebrin on the different stages of PRV infection. ( A ) Schematic timeline for the time-of-addition assays. ( B <t>)</t> <t>BTP−2</t> (600 nM) was co-incubated with PRV-QXX (MOI = 10) at 4 °C for 2 h to simulate virus adsorption under low-temperature conditions in PK−15 cells. Unbound viral particles were removed, viral genomic DNA was extracted, and the copy numbers of the viral genome were evaluated by qRT-PCR. ( C ) PK−15 cells were firstly incubated with PRV-QXX (MOI = 10) at 4 °C for 1 h to allow virus adsorption. Subsequently, unbound virus was removed, and medium containing 600 nM BTP−2 was added. Cells were treated at 37 °C for 1 h, and viral genome copy number was evaluated by qRT-PCR. ( D ) PK−15 cells were incubated with PRV−QXX (MOI = 1) at 4 °C for 1 h, and the inoculum was then removed and replaced with maintenance medium containing 2% FBS/DMEM. Cells were incubated at 37 °C for 12 h to permit viral entry and early replication events. Subsequently, BTP2 was added, and cells were incubated at 37 °C for another 12 h. Then, the virus was collected by being frozen and thawed three times, and the TCID 50 assay was performed to evaluate the virus titer. ( E ) PK−15 cells were treated as in ( D ): the supernatant was collected and TCID 50 was performed to evaluate the extracellular viral yield. ( F ) The sgCtrl and sgDBN cells were incubated with PRV-QXX at 4 °C for 2 h, cells were rinsed 3 times with ice-cold PBS and incubated with maintenance solution at 37 °C for 30 min, and then the PRV genome copy number was determined by RT-qPCR. ( G ) Cells were treated as in ( D ): viruses were harvested with three freeze–thaw cycles, and then the viral titers were analyzed by TCID 50 assay. ( H , I ) The sgCtrl and sgDBN cells were incubated with PRV−QXX at 37 °C for 1 h, cells were rinsed three times with PBS, and the medium was replaced with 2% FBS/DMEM to maintain growth for 12 h and 24 h. Then, the cells and supernatant were collected to analyze the intracellular and extracellular viral yield by the TCID 50 assay, respectively. Significance levels relative to the corresponding control were denoted as non-significant * p < 0.05, ** p < 0.01, and *** p < 0.001.
Ym 58483 Known Btp 2, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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btp-2 (# 203890 crac channel inhibitor btp2)  (Merck & Co)
90
Merck & Co btp-2 (# 203890 crac channel inhibitor btp2)
ER Ca 2+ release and SOCE contribute to the TRPML1-mediated Ca 2+ signals in hCMEC/D3 cells. (A, B) pre-incubation with CPA (10 μM, 30 min) or 2-APB (50 μM, 30 min) significantly reduced the ML-SA1-induced Ca 2+ response under 0Ca 2+ conditions. (C) Mean ± SEM of the amplitude of the Ca 2+ responses in cells under the designated treatments. **** indicates p < 0.0001 (Kruskal–Wallis one-way Anova test followed by Dunn’s post hoc test). (D) The Orai1 blockers, <t>BTP-2</t> (20 μM, 20 min) and Pyr6 (20 μM, 20 min), reduced the ML-SA1-evoked Ca 2+ response. (E) Mean ± SEM of the amplitude of Ca 2+ responses in cells under the designated treatments. Each drug totally inhibited the Ca 2+ response. **** indicates p < 0.0001 (Kruskal–Wallis one-way Anova test followed by the Dunn’s post hoc test).
Btp 2 (# 203890 Crac Channel Inhibitor Btp2), supplied by Merck & Co, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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4-methyl-4′-[3,5-bis(trifluoromethyl)-1h-pyrazol-1-yl]-1,2,3thiadiazole-5-carboxanilide (btp-2)  (Tocris)
90
Tocris 4-methyl-4′-[3,5-bis(trifluoromethyl)-1h-pyrazol-1-yl]-1,2,3thiadiazole-5-carboxanilide (btp-2)
ER Ca 2+ release and SOCE contribute to the TRPML1-mediated Ca 2+ signals in hCMEC/D3 cells. (A, B) pre-incubation with CPA (10 μM, 30 min) or 2-APB (50 μM, 30 min) significantly reduced the ML-SA1-induced Ca 2+ response under 0Ca 2+ conditions. (C) Mean ± SEM of the amplitude of the Ca 2+ responses in cells under the designated treatments. **** indicates p < 0.0001 (Kruskal–Wallis one-way Anova test followed by Dunn’s post hoc test). (D) The Orai1 blockers, <t>BTP-2</t> (20 μM, 20 min) and Pyr6 (20 μM, 20 min), reduced the ML-SA1-evoked Ca 2+ response. (E) Mean ± SEM of the amplitude of Ca 2+ responses in cells under the designated treatments. Each drug totally inhibited the Ca 2+ response. **** indicates p < 0.0001 (Kruskal–Wallis one-way Anova test followed by the Dunn’s post hoc test).
4 Methyl 4′ [3,5 Bis(Trifluoromethyl) 1h Pyrazol 1 Yl] 1,2,3thiadiazole 5 Carboxanilide (Btp 2), supplied by Tocris, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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btp-2  (Millipore)
90
Millipore btp-2
SOCE in ventricular myocytes is reduced by pharmacological inhibitors of TRPC and Orai channels. ( A ) Representative normalized fluorescence traces of CaTs at 1 Hz stimulation ( left ) and normalized fluorescence traces of SOCE ( right ) of an untreated control myocyte in the presence of DMSO ( a ) and of ventricular myocytes measured in the presence of 1 μM S66 ( b ), 3 μM <t>BTP-2</t> ( c ), 5 μM SKF96365 ( d ), or 1 μM S66 plus 5 μM SKF96365 ( e ). Cytosolic (black) and nuclear (red) traces are shown. ( B ) Averaged values of SOCE/CaT (%) for cytoplasm ( left ) and nucleus ( right ) of untreated control cells (ctl) and of cells treated with the various inhibitors, as indicated, depicted as scatter dot plots and box plots with medians. DMSO ctl: n = 35, N = 17; S66: n = 26, N = 4; BTP-2: n = 31, N = 5; SKF: n = 28, N = 5; S66/SKF: n = 20, N = 3; Kruskal–Wallis test with Dunn’s multiple comparisons test; p -values as indicated; *, p < 0.05; **, p < 0.01; ***, p < 0.001.
Btp 2, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Effects of Drebrin on the different stages of PRV infection. ( A ) Schematic timeline for the time-of-addition assays. ( B ) BTP−2 (600 nM) was co-incubated with PRV-QXX (MOI = 10) at 4 °C for 2 h to simulate virus adsorption under low-temperature conditions in PK−15 cells. Unbound viral particles were removed, viral genomic DNA was extracted, and the copy numbers of the viral genome were evaluated by qRT-PCR. ( C ) PK−15 cells were firstly incubated with PRV-QXX (MOI = 10) at 4 °C for 1 h to allow virus adsorption. Subsequently, unbound virus was removed, and medium containing 600 nM BTP−2 was added. Cells were treated at 37 °C for 1 h, and viral genome copy number was evaluated by qRT-PCR. ( D ) PK−15 cells were incubated with PRV−QXX (MOI = 1) at 4 °C for 1 h, and the inoculum was then removed and replaced with maintenance medium containing 2% FBS/DMEM. Cells were incubated at 37 °C for 12 h to permit viral entry and early replication events. Subsequently, BTP2 was added, and cells were incubated at 37 °C for another 12 h. Then, the virus was collected by being frozen and thawed three times, and the TCID 50 assay was performed to evaluate the virus titer. ( E ) PK−15 cells were treated as in ( D ): the supernatant was collected and TCID 50 was performed to evaluate the extracellular viral yield. ( F ) The sgCtrl and sgDBN cells were incubated with PRV-QXX at 4 °C for 2 h, cells were rinsed 3 times with ice-cold PBS and incubated with maintenance solution at 37 °C for 30 min, and then the PRV genome copy number was determined by RT-qPCR. ( G ) Cells were treated as in ( D ): viruses were harvested with three freeze–thaw cycles, and then the viral titers were analyzed by TCID 50 assay. ( H , I ) The sgCtrl and sgDBN cells were incubated with PRV−QXX at 37 °C for 1 h, cells were rinsed three times with PBS, and the medium was replaced with 2% FBS/DMEM to maintain growth for 12 h and 24 h. Then, the cells and supernatant were collected to analyze the intracellular and extracellular viral yield by the TCID 50 assay, respectively. Significance levels relative to the corresponding control were denoted as non-significant * p < 0.05, ** p < 0.01, and *** p < 0.001.

Journal: Microorganisms

Article Title: Drebrin Is Involved in the Life Cycle of Pseudorabies Virus by Regulating the Actin Cytoskeleton

doi: 10.3390/microorganisms13091969

Figure Lengend Snippet: Effects of Drebrin on the different stages of PRV infection. ( A ) Schematic timeline for the time-of-addition assays. ( B ) BTP−2 (600 nM) was co-incubated with PRV-QXX (MOI = 10) at 4 °C for 2 h to simulate virus adsorption under low-temperature conditions in PK−15 cells. Unbound viral particles were removed, viral genomic DNA was extracted, and the copy numbers of the viral genome were evaluated by qRT-PCR. ( C ) PK−15 cells were firstly incubated with PRV-QXX (MOI = 10) at 4 °C for 1 h to allow virus adsorption. Subsequently, unbound virus was removed, and medium containing 600 nM BTP−2 was added. Cells were treated at 37 °C for 1 h, and viral genome copy number was evaluated by qRT-PCR. ( D ) PK−15 cells were incubated with PRV−QXX (MOI = 1) at 4 °C for 1 h, and the inoculum was then removed and replaced with maintenance medium containing 2% FBS/DMEM. Cells were incubated at 37 °C for 12 h to permit viral entry and early replication events. Subsequently, BTP2 was added, and cells were incubated at 37 °C for another 12 h. Then, the virus was collected by being frozen and thawed three times, and the TCID 50 assay was performed to evaluate the virus titer. ( E ) PK−15 cells were treated as in ( D ): the supernatant was collected and TCID 50 was performed to evaluate the extracellular viral yield. ( F ) The sgCtrl and sgDBN cells were incubated with PRV-QXX at 4 °C for 2 h, cells were rinsed 3 times with ice-cold PBS and incubated with maintenance solution at 37 °C for 30 min, and then the PRV genome copy number was determined by RT-qPCR. ( G ) Cells were treated as in ( D ): viruses were harvested with three freeze–thaw cycles, and then the viral titers were analyzed by TCID 50 assay. ( H , I ) The sgCtrl and sgDBN cells were incubated with PRV−QXX at 37 °C for 1 h, cells were rinsed three times with PBS, and the medium was replaced with 2% FBS/DMEM to maintain growth for 12 h and 24 h. Then, the cells and supernatant were collected to analyze the intracellular and extracellular viral yield by the TCID 50 assay, respectively. Significance levels relative to the corresponding control were denoted as non-significant * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: BTP−2 (HY-100831) and Puromycin (P8230) were obtained from MedChemExpress (South Brunswick Township, NJ, USA) and Solarbio (Beijing, China), respectively.

Techniques: Infection, Incubation, Virus, Adsorption, Quantitative RT-PCR, Control

ER Ca 2+ release and SOCE contribute to the TRPML1-mediated Ca 2+ signals in hCMEC/D3 cells. (A, B) pre-incubation with CPA (10 μM, 30 min) or 2-APB (50 μM, 30 min) significantly reduced the ML-SA1-induced Ca 2+ response under 0Ca 2+ conditions. (C) Mean ± SEM of the amplitude of the Ca 2+ responses in cells under the designated treatments. **** indicates p < 0.0001 (Kruskal–Wallis one-way Anova test followed by Dunn’s post hoc test). (D) The Orai1 blockers, BTP-2 (20 μM, 20 min) and Pyr6 (20 μM, 20 min), reduced the ML-SA1-evoked Ca 2+ response. (E) Mean ± SEM of the amplitude of Ca 2+ responses in cells under the designated treatments. Each drug totally inhibited the Ca 2+ response. **** indicates p < 0.0001 (Kruskal–Wallis one-way Anova test followed by the Dunn’s post hoc test).

Journal: Frontiers in Physiology

Article Title: Lysosomal TRPML1 triggers global Ca 2+ signals and nitric oxide release in human cerebrovascular endothelial cells

doi: 10.3389/fphys.2024.1426783

Figure Lengend Snippet: ER Ca 2+ release and SOCE contribute to the TRPML1-mediated Ca 2+ signals in hCMEC/D3 cells. (A, B) pre-incubation with CPA (10 μM, 30 min) or 2-APB (50 μM, 30 min) significantly reduced the ML-SA1-induced Ca 2+ response under 0Ca 2+ conditions. (C) Mean ± SEM of the amplitude of the Ca 2+ responses in cells under the designated treatments. **** indicates p < 0.0001 (Kruskal–Wallis one-way Anova test followed by Dunn’s post hoc test). (D) The Orai1 blockers, BTP-2 (20 μM, 20 min) and Pyr6 (20 μM, 20 min), reduced the ML-SA1-evoked Ca 2+ response. (E) Mean ± SEM of the amplitude of Ca 2+ responses in cells under the designated treatments. Each drug totally inhibited the Ca 2+ response. **** indicates p < 0.0001 (Kruskal–Wallis one-way Anova test followed by the Dunn’s post hoc test).

Article Snippet: ML-SAI (# SML0627), ML-SI3 (GW405833 hydrochloride; # G1421), CPA (Ciclo piazonic acid; # C1530), 2-APB (# D9754), BTP-2 (# 203890 CRAC channel inhibitor BTP2), Pyr6 (# SML1241), were purchased from Merck GKaA (Darmstadt, Germany).

Techniques: Incubation

SOCE in ventricular myocytes is reduced by pharmacological inhibitors of TRPC and Orai channels. ( A ) Representative normalized fluorescence traces of CaTs at 1 Hz stimulation ( left ) and normalized fluorescence traces of SOCE ( right ) of an untreated control myocyte in the presence of DMSO ( a ) and of ventricular myocytes measured in the presence of 1 μM S66 ( b ), 3 μM BTP-2 ( c ), 5 μM SKF96365 ( d ), or 1 μM S66 plus 5 μM SKF96365 ( e ). Cytosolic (black) and nuclear (red) traces are shown. ( B ) Averaged values of SOCE/CaT (%) for cytoplasm ( left ) and nucleus ( right ) of untreated control cells (ctl) and of cells treated with the various inhibitors, as indicated, depicted as scatter dot plots and box plots with medians. DMSO ctl: n = 35, N = 17; S66: n = 26, N = 4; BTP-2: n = 31, N = 5; SKF: n = 28, N = 5; S66/SKF: n = 20, N = 3; Kruskal–Wallis test with Dunn’s multiple comparisons test; p -values as indicated; *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Journal: Cells

Article Title: Store-Operated Calcium Entry Increases Nuclear Calcium in Adult Rat Atrial and Ventricular Cardiomyocytes

doi: 10.3390/cells12232690

Figure Lengend Snippet: SOCE in ventricular myocytes is reduced by pharmacological inhibitors of TRPC and Orai channels. ( A ) Representative normalized fluorescence traces of CaTs at 1 Hz stimulation ( left ) and normalized fluorescence traces of SOCE ( right ) of an untreated control myocyte in the presence of DMSO ( a ) and of ventricular myocytes measured in the presence of 1 μM S66 ( b ), 3 μM BTP-2 ( c ), 5 μM SKF96365 ( d ), or 1 μM S66 plus 5 μM SKF96365 ( e ). Cytosolic (black) and nuclear (red) traces are shown. ( B ) Averaged values of SOCE/CaT (%) for cytoplasm ( left ) and nucleus ( right ) of untreated control cells (ctl) and of cells treated with the various inhibitors, as indicated, depicted as scatter dot plots and box plots with medians. DMSO ctl: n = 35, N = 17; S66: n = 26, N = 4; BTP-2: n = 31, N = 5; SKF: n = 28, N = 5; S66/SKF: n = 20, N = 3; Kruskal–Wallis test with Dunn’s multiple comparisons test; p -values as indicated; *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: Following store depletion and another minute of resting, the extracellular solution was switched to a 2 mM Ca solution containing (mM) 130 NaCl, 5.4 KCl, 2 CaCl 2 , 0.5 MgCl 2 , 10 HEPES, and 10 glucose, adjusted to pH 7.4, and cytosolic and nuclear fluorescence increases were recorded for 90 s. The TRPC channel blocker SKF96365 (SKF, Cayman Chemical) and the Orai/CRAC channel inhibitors Synta 66 (S66, Sigma-Aldrich, Taufkirchen, Germany) and BTP-2 (Sigma-Aldrich) were used to study the impact of the respective channels on SOCE in cardiomyocytes.

Techniques: Fluorescence

SOCE in atrial myocytes is reduced by pharmacological inhibitors of TRPC and Orai channels. ( A ) Representative normalized fluorescence traces of CaTs at 0.5 Hz stimulation ( left ) and normalized fluorescence traces of SOCE ( right ) of an untreated control atrial myocyte in the presence of DMSO ( a ) and of atrial myocytes measured in the presence of 1 μM S66 ( b ), 3 μM BTP-2 ( c ), 5 μM SKF96365 ( d ), or 1 μM S66 plus 5 μM SKF96365 ( e ). Cytosolic (black) and nuclear (red) traces are shown. ( B ) Averaged values of SOCE/CaT (%) for cytoplasm ( left ) and nucleus ( right ) of untreated control cells (ctl) and of cells treated with the various inhibitors, as indicated, depicted as scatter dot plots and box plots with medians. Kruskal–Wallis test with Dunn’s multiple comparisons test; DMSO ctl: n = 41, N = 17; S66: n = 27, N = 5; BTP-2: n = 29, N = 4; SKF: n = 32, N = 4; S66/SKF: n = 28, N = 7; p -values as indicated; ***, p < 0.001.

Journal: Cells

Article Title: Store-Operated Calcium Entry Increases Nuclear Calcium in Adult Rat Atrial and Ventricular Cardiomyocytes

doi: 10.3390/cells12232690

Figure Lengend Snippet: SOCE in atrial myocytes is reduced by pharmacological inhibitors of TRPC and Orai channels. ( A ) Representative normalized fluorescence traces of CaTs at 0.5 Hz stimulation ( left ) and normalized fluorescence traces of SOCE ( right ) of an untreated control atrial myocyte in the presence of DMSO ( a ) and of atrial myocytes measured in the presence of 1 μM S66 ( b ), 3 μM BTP-2 ( c ), 5 μM SKF96365 ( d ), or 1 μM S66 plus 5 μM SKF96365 ( e ). Cytosolic (black) and nuclear (red) traces are shown. ( B ) Averaged values of SOCE/CaT (%) for cytoplasm ( left ) and nucleus ( right ) of untreated control cells (ctl) and of cells treated with the various inhibitors, as indicated, depicted as scatter dot plots and box plots with medians. Kruskal–Wallis test with Dunn’s multiple comparisons test; DMSO ctl: n = 41, N = 17; S66: n = 27, N = 5; BTP-2: n = 29, N = 4; SKF: n = 32, N = 4; S66/SKF: n = 28, N = 7; p -values as indicated; ***, p < 0.001.

Article Snippet: Following store depletion and another minute of resting, the extracellular solution was switched to a 2 mM Ca solution containing (mM) 130 NaCl, 5.4 KCl, 2 CaCl 2 , 0.5 MgCl 2 , 10 HEPES, and 10 glucose, adjusted to pH 7.4, and cytosolic and nuclear fluorescence increases were recorded for 90 s. The TRPC channel blocker SKF96365 (SKF, Cayman Chemical) and the Orai/CRAC channel inhibitors Synta 66 (S66, Sigma-Aldrich, Taufkirchen, Germany) and BTP-2 (Sigma-Aldrich) were used to study the impact of the respective channels on SOCE in cardiomyocytes.

Techniques: Fluorescence