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1 2 dioctanoyl phosphatidylinositol 4 5 bisphosphate sodium salt  (Cayman Chemical)

 
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    Cayman Chemical 1 2 dioctanoyl phosphatidylinositol 4 5 bisphosphate sodium salt
    ( A ) Schematic diagram showing the ATP-dependent synthesis steps leading to the production of PIP 2 . ( B ) Average maximum outward TRPV4 current induced by 100 nM GSK101, recorded in cECs at 100 mV using the conventional whole-cell configuration. cECs dialyzed with 1 mM Mg-ATP were treated for ~10 min with wortmannin (0.1, 50 µM), PIK93 (0.3 µM), PAO (30 µM) or LY294002 (10, 300 µM), or were left untreated (control). A minimum duration of 10–15 min after the application of GSK101 was allowed for outward TRPV4 current to develop in each cEC. Data are means ± SEM (**p<0.01, *p<0.05 vs. control Mg-ATP, one-way ANOVA followed by Dunnett’s multiple comparisons test; n = 6–27). ( C ) Traces of current-voltage relationship obtained from a cEC dialyzed with 10 µM <t>diC8-PIP</t> 2 and 0 mM Mg-ATP using a voltage ramp (−100 to 100 mV) before and after (green) the application of GSK101 (100 nM) and RuR (1 µM). The dotted gray trace is a representative GSK101-induced current recorded from a control cEC dialyzed with 0 µM diC8-PIP 2 and 0 mM Mg-ATP. ( D ) Summary data showing GSK101 (100 nM)-induced currents at 100 mV in cECs dialyzed with different concentrations of diC8-PIP 2 (10, 20, 50 µM) or 0 µM phosphoinositide (control). The pipette solution lacked Mg-ATP in all groups. GSK101-evoked outward currents developed over ~5 min. Data are presented as means ± SEM (*p<0.05, * * P <0.01, one-way ANOVA followed by Dunnett’s multiple comparisons test; n = 10–18). ( E, F ) Representative trace ( E ) and summary data showing GSK101-induced currents in cECs dialyzed with 1 mM Mg-ATP and poly-L-lysine (3 µg/ml). A duration of 10 min was allowed after the application of GSK101 for outward TRPV4 current to develop in each cEC. Data in F are presented as means ± SEM (**p<0.01, unpaired Student’s t-test; n = 8–18). 10.7554/eLife.38689.017 Figure 3—source data 1. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.018 Figure 3—source data 2. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.019 Figure 3—source data 3. Numerical data that were used to generate the chart in .
    1 2 Dioctanoyl Phosphatidylinositol 4 5 Bisphosphate Sodium Salt, supplied by Cayman Chemical, 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/1 2 dioctanoyl phosphatidylinositol 4 5 bisphosphate sodium salt/product/Cayman Chemical
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    1 2 dioctanoyl phosphatidylinositol 4 5 bisphosphate sodium salt - by Bioz Stars, 2024-12
    86/100 stars

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    1) Product Images from "PIP 2 depletion promotes TRPV4 channel activity in mouse brain capillary endothelial cells"

    Article Title: PIP 2 depletion promotes TRPV4 channel activity in mouse brain capillary endothelial cells

    Journal: eLife

    doi: 10.7554/eLife.38689

    ( A ) Schematic diagram showing the ATP-dependent synthesis steps leading to the production of PIP 2 . ( B ) Average maximum outward TRPV4 current induced by 100 nM GSK101, recorded in cECs at 100 mV using the conventional whole-cell configuration. cECs dialyzed with 1 mM Mg-ATP were treated for ~10 min with wortmannin (0.1, 50 µM), PIK93 (0.3 µM), PAO (30 µM) or LY294002 (10, 300 µM), or were left untreated (control). A minimum duration of 10–15 min after the application of GSK101 was allowed for outward TRPV4 current to develop in each cEC. Data are means ± SEM (**p<0.01, *p<0.05 vs. control Mg-ATP, one-way ANOVA followed by Dunnett’s multiple comparisons test; n = 6–27). ( C ) Traces of current-voltage relationship obtained from a cEC dialyzed with 10 µM diC8-PIP 2 and 0 mM Mg-ATP using a voltage ramp (−100 to 100 mV) before and after (green) the application of GSK101 (100 nM) and RuR (1 µM). The dotted gray trace is a representative GSK101-induced current recorded from a control cEC dialyzed with 0 µM diC8-PIP 2 and 0 mM Mg-ATP. ( D ) Summary data showing GSK101 (100 nM)-induced currents at 100 mV in cECs dialyzed with different concentrations of diC8-PIP 2 (10, 20, 50 µM) or 0 µM phosphoinositide (control). The pipette solution lacked Mg-ATP in all groups. GSK101-evoked outward currents developed over ~5 min. Data are presented as means ± SEM (*p<0.05, * * P <0.01, one-way ANOVA followed by Dunnett’s multiple comparisons test; n = 10–18). ( E, F ) Representative trace ( E ) and summary data showing GSK101-induced currents in cECs dialyzed with 1 mM Mg-ATP and poly-L-lysine (3 µg/ml). A duration of 10 min was allowed after the application of GSK101 for outward TRPV4 current to develop in each cEC. Data in F are presented as means ± SEM (**p<0.01, unpaired Student’s t-test; n = 8–18). 10.7554/eLife.38689.017 Figure 3—source data 1. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.018 Figure 3—source data 2. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.019 Figure 3—source data 3. Numerical data that were used to generate the chart in .
    Figure Legend Snippet: ( A ) Schematic diagram showing the ATP-dependent synthesis steps leading to the production of PIP 2 . ( B ) Average maximum outward TRPV4 current induced by 100 nM GSK101, recorded in cECs at 100 mV using the conventional whole-cell configuration. cECs dialyzed with 1 mM Mg-ATP were treated for ~10 min with wortmannin (0.1, 50 µM), PIK93 (0.3 µM), PAO (30 µM) or LY294002 (10, 300 µM), or were left untreated (control). A minimum duration of 10–15 min after the application of GSK101 was allowed for outward TRPV4 current to develop in each cEC. Data are means ± SEM (**p<0.01, *p<0.05 vs. control Mg-ATP, one-way ANOVA followed by Dunnett’s multiple comparisons test; n = 6–27). ( C ) Traces of current-voltage relationship obtained from a cEC dialyzed with 10 µM diC8-PIP 2 and 0 mM Mg-ATP using a voltage ramp (−100 to 100 mV) before and after (green) the application of GSK101 (100 nM) and RuR (1 µM). The dotted gray trace is a representative GSK101-induced current recorded from a control cEC dialyzed with 0 µM diC8-PIP 2 and 0 mM Mg-ATP. ( D ) Summary data showing GSK101 (100 nM)-induced currents at 100 mV in cECs dialyzed with different concentrations of diC8-PIP 2 (10, 20, 50 µM) or 0 µM phosphoinositide (control). The pipette solution lacked Mg-ATP in all groups. GSK101-evoked outward currents developed over ~5 min. Data are presented as means ± SEM (*p<0.05, * * P <0.01, one-way ANOVA followed by Dunnett’s multiple comparisons test; n = 10–18). ( E, F ) Representative trace ( E ) and summary data showing GSK101-induced currents in cECs dialyzed with 1 mM Mg-ATP and poly-L-lysine (3 µg/ml). A duration of 10 min was allowed after the application of GSK101 for outward TRPV4 current to develop in each cEC. Data in F are presented as means ± SEM (**p<0.01, unpaired Student’s t-test; n = 8–18). 10.7554/eLife.38689.017 Figure 3—source data 1. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.018 Figure 3—source data 2. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.019 Figure 3—source data 3. Numerical data that were used to generate the chart in .

    Techniques Used: Transferring

    ( A ) Chemical structures of short (dioctanoyl, diC8-PIP 2 ) and long (dipalmitoyl, diC16-PIP 2 ) acyl chain PIP 2 salts used in this study. ( B ) Current-voltage relationship represents currents obtained from a cEC dialyzed with 10 µM diC16-PIP 2 and 0 mM Mg-ATP before (light green) and after (dark green) the application of GSK101 (100 nM) and RuR (1 µM), recorded using a voltage ramp (−100 to 100 mV). Outward current developed over 5 min. The dotted gray trace is a representative GSK101-induced current recorded from a control cECs dialyzed with 0 mM diC16-PIP 2 and 0 mM Mg-ATP. ( C ) Summary data showing GSK101 (100 nM)-induced currents at 100 mV in cECs dialyzed with diC16-PIP 2 (10 µM) or 0 µM phosphoinositide (control). The pipette solution lacked Mg-ATP in both groups. Data are presented as means ± SEM (*p<0.05, unpaired Student’s t-test; n = 3–4 cECs).
    Figure Legend Snippet: ( A ) Chemical structures of short (dioctanoyl, diC8-PIP 2 ) and long (dipalmitoyl, diC16-PIP 2 ) acyl chain PIP 2 salts used in this study. ( B ) Current-voltage relationship represents currents obtained from a cEC dialyzed with 10 µM diC16-PIP 2 and 0 mM Mg-ATP before (light green) and after (dark green) the application of GSK101 (100 nM) and RuR (1 µM), recorded using a voltage ramp (−100 to 100 mV). Outward current developed over 5 min. The dotted gray trace is a representative GSK101-induced current recorded from a control cECs dialyzed with 0 mM diC16-PIP 2 and 0 mM Mg-ATP. ( C ) Summary data showing GSK101 (100 nM)-induced currents at 100 mV in cECs dialyzed with diC16-PIP 2 (10 µM) or 0 µM phosphoinositide (control). The pipette solution lacked Mg-ATP in both groups. Data are presented as means ± SEM (*p<0.05, unpaired Student’s t-test; n = 3–4 cECs).

    Techniques Used: Transferring

    ( A ) Representative current-voltage plots obtained from a cEC dialyzed with 1 mM Mg-ATP and treated consecutively with GSK101 (100 nM) and RuR (1 µM) followed by 2 µM PGE 2 . ( B ) Summary individual-value plot of GSK101-induced TRPV4 currents at 100 mV in cECs dialyzed with 1 mM Mg-ATP in the absence (black; n = 27) and presence (orange; n = 15) of 2 µM PGE 2 . Incubation of cECs with PGE 2 lasted ~15 min. Data in B are means (column bars) ± SEM (error bars, ****p<0.0001, unpaired Student’s t-test). ( C ) Top: Schematic diagram showing the G q PCR-dependent hydrolysis of PIP 2 and the interventions used to test different components of the proposed pathway. Bottom: Summary data showing GSK101 (100 nM)-induced currents recorded at 100 mV in cECs dialyzed with 1 mM Mg-ATP. Currents were recorded in the absence and presence of 2 µM PGE 2 (orange shading), with or without (control) the indicated interventions. Concentrations (and application method): HC-067047, 1 µM (bath); diC8-PIP 2 , 10 µM (pipette), U73122, 10 µM (bath); U73343, 10 µM (bath); AH6809, 10 µM (bath); SC51322, 1 µM (bath); calphostin C, 0.5 µM (bath); Gö6976, 1 µM (bath); CPA, 30 µM (bath); BAPTA, 5.4 mM (pipette). For bath application, pharmacological agents were added 10–15 min before PGE 2 application. 10.7554/eLife.38689.025 Figure 5—source data 1. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.026 Figure 5—source data 2. Numerical data that were used to generate the chart in .
    Figure Legend Snippet: ( A ) Representative current-voltage plots obtained from a cEC dialyzed with 1 mM Mg-ATP and treated consecutively with GSK101 (100 nM) and RuR (1 µM) followed by 2 µM PGE 2 . ( B ) Summary individual-value plot of GSK101-induced TRPV4 currents at 100 mV in cECs dialyzed with 1 mM Mg-ATP in the absence (black; n = 27) and presence (orange; n = 15) of 2 µM PGE 2 . Incubation of cECs with PGE 2 lasted ~15 min. Data in B are means (column bars) ± SEM (error bars, ****p<0.0001, unpaired Student’s t-test). ( C ) Top: Schematic diagram showing the G q PCR-dependent hydrolysis of PIP 2 and the interventions used to test different components of the proposed pathway. Bottom: Summary data showing GSK101 (100 nM)-induced currents recorded at 100 mV in cECs dialyzed with 1 mM Mg-ATP. Currents were recorded in the absence and presence of 2 µM PGE 2 (orange shading), with or without (control) the indicated interventions. Concentrations (and application method): HC-067047, 1 µM (bath); diC8-PIP 2 , 10 µM (pipette), U73122, 10 µM (bath); U73343, 10 µM (bath); AH6809, 10 µM (bath); SC51322, 1 µM (bath); calphostin C, 0.5 µM (bath); Gö6976, 1 µM (bath); CPA, 30 µM (bath); BAPTA, 5.4 mM (pipette). For bath application, pharmacological agents were added 10–15 min before PGE 2 application. 10.7554/eLife.38689.025 Figure 5—source data 1. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.026 Figure 5—source data 2. Numerical data that were used to generate the chart in .

    Techniques Used: Incubation, Transferring

    ( A ) Current-voltage relationship recorded in a cEC dialyzed with 1 mM Mg-ATP using voltage ramps (−100 to 100 mV). The circled numbers indicated the sequential and cumulative application of GSK101 (100 nM)+RuR (1 µM) followed by 2 µM PGE 2 and lastly, the TRPV4 blocker HC-067047 (1 µM). ( B ) Representative scatter plots of peak current amplitudes (at 100 mV) in three cECs dialyzed with 1 mM Mg-ATP and treated with 100 nM GSK101 followed by 2 µM PGE 2 (or PGE 2 first followed by GSK101). Note that PGE 2 -induced activation of TRPV4 current plateaued after ~13–15 min. ( C ) Scatter plots of normalized GSK101-induced outward currents (at 100 mV), normalized to the current at zero time, I 0 (when 2 µM PGE 2 was applied), in two cECs, one dialyzed with 1 mM Mg-ATP (orange symbols) and the other dialyzed with 1 mM Mg-ATP +10 µM diC8-PIP 2 (black symbols). Currents normalized to I 0 are presented before and after the application of PGE 2 for 35 min.
    Figure Legend Snippet: ( A ) Current-voltage relationship recorded in a cEC dialyzed with 1 mM Mg-ATP using voltage ramps (−100 to 100 mV). The circled numbers indicated the sequential and cumulative application of GSK101 (100 nM)+RuR (1 µM) followed by 2 µM PGE 2 and lastly, the TRPV4 blocker HC-067047 (1 µM). ( B ) Representative scatter plots of peak current amplitudes (at 100 mV) in three cECs dialyzed with 1 mM Mg-ATP and treated with 100 nM GSK101 followed by 2 µM PGE 2 (or PGE 2 first followed by GSK101). Note that PGE 2 -induced activation of TRPV4 current plateaued after ~13–15 min. ( C ) Scatter plots of normalized GSK101-induced outward currents (at 100 mV), normalized to the current at zero time, I 0 (when 2 µM PGE 2 was applied), in two cECs, one dialyzed with 1 mM Mg-ATP (orange symbols) and the other dialyzed with 1 mM Mg-ATP +10 µM diC8-PIP 2 (black symbols). Currents normalized to I 0 are presented before and after the application of PGE 2 for 35 min.

    Techniques Used: Activation Assay

    ( A ) Current-voltage relationship represent currents obtained from a cEC dialyzed with 0 µM diC8-PIP 2 and 0 mM Mg-ATP before (control) and after the application of 11,12-EET (1 µM) and RuR (1 µM), recorded using 300 ms voltage ramps (−100 to 100 mV). ( B ) Summary data showing 11,12-EET-induced outward currents at 100 mV in dialyzed cECs based on experiments in A (n = 6 cECs).
    Figure Legend Snippet: ( A ) Current-voltage relationship represent currents obtained from a cEC dialyzed with 0 µM diC8-PIP 2 and 0 mM Mg-ATP before (control) and after the application of 11,12-EET (1 µM) and RuR (1 µM), recorded using 300 ms voltage ramps (−100 to 100 mV). ( B ) Summary data showing 11,12-EET-induced outward currents at 100 mV in dialyzed cECs based on experiments in A (n = 6 cECs).

    Techniques Used:



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    1 2 dioctanoyl phosphatidylinositol 4 5 bisphosphate sodium salt  (Cayman Chemical)
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    Cayman Chemical 1 2 dioctanoyl phosphatidylinositol 4 5 bisphosphate sodium salt
    ( A ) Schematic diagram showing the ATP-dependent synthesis steps leading to the production of PIP 2 . ( B ) Average maximum outward TRPV4 current induced by 100 nM GSK101, recorded in cECs at 100 mV using the conventional whole-cell configuration. cECs dialyzed with 1 mM Mg-ATP were treated for ~10 min with wortmannin (0.1, 50 µM), PIK93 (0.3 µM), PAO (30 µM) or LY294002 (10, 300 µM), or were left untreated (control). A minimum duration of 10–15 min after the application of GSK101 was allowed for outward TRPV4 current to develop in each cEC. Data are means ± SEM (**p<0.01, *p<0.05 vs. control Mg-ATP, one-way ANOVA followed by Dunnett’s multiple comparisons test; n = 6–27). ( C ) Traces of current-voltage relationship obtained from a cEC dialyzed with 10 µM <t>diC8-PIP</t> 2 and 0 mM Mg-ATP using a voltage ramp (−100 to 100 mV) before and after (green) the application of GSK101 (100 nM) and RuR (1 µM). The dotted gray trace is a representative GSK101-induced current recorded from a control cEC dialyzed with 0 µM diC8-PIP 2 and 0 mM Mg-ATP. ( D ) Summary data showing GSK101 (100 nM)-induced currents at 100 mV in cECs dialyzed with different concentrations of diC8-PIP 2 (10, 20, 50 µM) or 0 µM phosphoinositide (control). The pipette solution lacked Mg-ATP in all groups. GSK101-evoked outward currents developed over ~5 min. Data are presented as means ± SEM (*p<0.05, * * P <0.01, one-way ANOVA followed by Dunnett’s multiple comparisons test; n = 10–18). ( E, F ) Representative trace ( E ) and summary data showing GSK101-induced currents in cECs dialyzed with 1 mM Mg-ATP and poly-L-lysine (3 µg/ml). A duration of 10 min was allowed after the application of GSK101 for outward TRPV4 current to develop in each cEC. Data in F are presented as means ± SEM (**p<0.01, unpaired Student’s t-test; n = 8–18). 10.7554/eLife.38689.017 Figure 3—source data 1. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.018 Figure 3—source data 2. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.019 Figure 3—source data 3. Numerical data that were used to generate the chart in .
    1 2 Dioctanoyl Phosphatidylinositol 4 5 Bisphosphate Sodium Salt, supplied by Cayman Chemical, 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/1 2 dioctanoyl phosphatidylinositol 4 5 bisphosphate sodium salt/product/Cayman Chemical
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    1 2 dioctanoyl phosphatidylinositol 4 5 bisphosphate sodium salt - by Bioz Stars, 2024-12
    86/100 stars
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    ( A ) Schematic diagram showing the ATP-dependent synthesis steps leading to the production of PIP 2 . ( B ) Average maximum outward TRPV4 current induced by 100 nM GSK101, recorded in cECs at 100 mV using the conventional whole-cell configuration. cECs dialyzed with 1 mM Mg-ATP were treated for ~10 min with wortmannin (0.1, 50 µM), PIK93 (0.3 µM), PAO (30 µM) or LY294002 (10, 300 µM), or were left untreated (control). A minimum duration of 10–15 min after the application of GSK101 was allowed for outward TRPV4 current to develop in each cEC. Data are means ± SEM (**p<0.01, *p<0.05 vs. control Mg-ATP, one-way ANOVA followed by Dunnett’s multiple comparisons test; n = 6–27). ( C ) Traces of current-voltage relationship obtained from a cEC dialyzed with 10 µM diC8-PIP 2 and 0 mM Mg-ATP using a voltage ramp (−100 to 100 mV) before and after (green) the application of GSK101 (100 nM) and RuR (1 µM). The dotted gray trace is a representative GSK101-induced current recorded from a control cEC dialyzed with 0 µM diC8-PIP 2 and 0 mM Mg-ATP. ( D ) Summary data showing GSK101 (100 nM)-induced currents at 100 mV in cECs dialyzed with different concentrations of diC8-PIP 2 (10, 20, 50 µM) or 0 µM phosphoinositide (control). The pipette solution lacked Mg-ATP in all groups. GSK101-evoked outward currents developed over ~5 min. Data are presented as means ± SEM (*p<0.05, * * P <0.01, one-way ANOVA followed by Dunnett’s multiple comparisons test; n = 10–18). ( E, F ) Representative trace ( E ) and summary data showing GSK101-induced currents in cECs dialyzed with 1 mM Mg-ATP and poly-L-lysine (3 µg/ml). A duration of 10 min was allowed after the application of GSK101 for outward TRPV4 current to develop in each cEC. Data in F are presented as means ± SEM (**p<0.01, unpaired Student’s t-test; n = 8–18). 10.7554/eLife.38689.017 Figure 3—source data 1. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.018 Figure 3—source data 2. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.019 Figure 3—source data 3. Numerical data that were used to generate the chart in .

    Journal: eLife

    Article Title: PIP 2 depletion promotes TRPV4 channel activity in mouse brain capillary endothelial cells

    doi: 10.7554/eLife.38689

    Figure Lengend Snippet: ( A ) Schematic diagram showing the ATP-dependent synthesis steps leading to the production of PIP 2 . ( B ) Average maximum outward TRPV4 current induced by 100 nM GSK101, recorded in cECs at 100 mV using the conventional whole-cell configuration. cECs dialyzed with 1 mM Mg-ATP were treated for ~10 min with wortmannin (0.1, 50 µM), PIK93 (0.3 µM), PAO (30 µM) or LY294002 (10, 300 µM), or were left untreated (control). A minimum duration of 10–15 min after the application of GSK101 was allowed for outward TRPV4 current to develop in each cEC. Data are means ± SEM (**p<0.01, *p<0.05 vs. control Mg-ATP, one-way ANOVA followed by Dunnett’s multiple comparisons test; n = 6–27). ( C ) Traces of current-voltage relationship obtained from a cEC dialyzed with 10 µM diC8-PIP 2 and 0 mM Mg-ATP using a voltage ramp (−100 to 100 mV) before and after (green) the application of GSK101 (100 nM) and RuR (1 µM). The dotted gray trace is a representative GSK101-induced current recorded from a control cEC dialyzed with 0 µM diC8-PIP 2 and 0 mM Mg-ATP. ( D ) Summary data showing GSK101 (100 nM)-induced currents at 100 mV in cECs dialyzed with different concentrations of diC8-PIP 2 (10, 20, 50 µM) or 0 µM phosphoinositide (control). The pipette solution lacked Mg-ATP in all groups. GSK101-evoked outward currents developed over ~5 min. Data are presented as means ± SEM (*p<0.05, * * P <0.01, one-way ANOVA followed by Dunnett’s multiple comparisons test; n = 10–18). ( E, F ) Representative trace ( E ) and summary data showing GSK101-induced currents in cECs dialyzed with 1 mM Mg-ATP and poly-L-lysine (3 µg/ml). A duration of 10 min was allowed after the application of GSK101 for outward TRPV4 current to develop in each cEC. Data in F are presented as means ± SEM (**p<0.01, unpaired Student’s t-test; n = 8–18). 10.7554/eLife.38689.017 Figure 3—source data 1. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.018 Figure 3—source data 2. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.019 Figure 3—source data 3. Numerical data that were used to generate the chart in .

    Article Snippet: 1,2-Dioctanoyl phosphatidylinositol 4,5-bisphosphate sodium salt (diC8-PIP 2 ) and 1,2-dipalmitoyl phosphatidylinositol 4,5-bisphosphate sodium salt (diC16-PIP 2 ) were purchased from Cayman Chemical (USA).

    Techniques: Transferring

    ( A ) Chemical structures of short (dioctanoyl, diC8-PIP 2 ) and long (dipalmitoyl, diC16-PIP 2 ) acyl chain PIP 2 salts used in this study. ( B ) Current-voltage relationship represents currents obtained from a cEC dialyzed with 10 µM diC16-PIP 2 and 0 mM Mg-ATP before (light green) and after (dark green) the application of GSK101 (100 nM) and RuR (1 µM), recorded using a voltage ramp (−100 to 100 mV). Outward current developed over 5 min. The dotted gray trace is a representative GSK101-induced current recorded from a control cECs dialyzed with 0 mM diC16-PIP 2 and 0 mM Mg-ATP. ( C ) Summary data showing GSK101 (100 nM)-induced currents at 100 mV in cECs dialyzed with diC16-PIP 2 (10 µM) or 0 µM phosphoinositide (control). The pipette solution lacked Mg-ATP in both groups. Data are presented as means ± SEM (*p<0.05, unpaired Student’s t-test; n = 3–4 cECs).

    Journal: eLife

    Article Title: PIP 2 depletion promotes TRPV4 channel activity in mouse brain capillary endothelial cells

    doi: 10.7554/eLife.38689

    Figure Lengend Snippet: ( A ) Chemical structures of short (dioctanoyl, diC8-PIP 2 ) and long (dipalmitoyl, diC16-PIP 2 ) acyl chain PIP 2 salts used in this study. ( B ) Current-voltage relationship represents currents obtained from a cEC dialyzed with 10 µM diC16-PIP 2 and 0 mM Mg-ATP before (light green) and after (dark green) the application of GSK101 (100 nM) and RuR (1 µM), recorded using a voltage ramp (−100 to 100 mV). Outward current developed over 5 min. The dotted gray trace is a representative GSK101-induced current recorded from a control cECs dialyzed with 0 mM diC16-PIP 2 and 0 mM Mg-ATP. ( C ) Summary data showing GSK101 (100 nM)-induced currents at 100 mV in cECs dialyzed with diC16-PIP 2 (10 µM) or 0 µM phosphoinositide (control). The pipette solution lacked Mg-ATP in both groups. Data are presented as means ± SEM (*p<0.05, unpaired Student’s t-test; n = 3–4 cECs).

    Article Snippet: 1,2-Dioctanoyl phosphatidylinositol 4,5-bisphosphate sodium salt (diC8-PIP 2 ) and 1,2-dipalmitoyl phosphatidylinositol 4,5-bisphosphate sodium salt (diC16-PIP 2 ) were purchased from Cayman Chemical (USA).

    Techniques: Transferring

    ( A ) Representative current-voltage plots obtained from a cEC dialyzed with 1 mM Mg-ATP and treated consecutively with GSK101 (100 nM) and RuR (1 µM) followed by 2 µM PGE 2 . ( B ) Summary individual-value plot of GSK101-induced TRPV4 currents at 100 mV in cECs dialyzed with 1 mM Mg-ATP in the absence (black; n = 27) and presence (orange; n = 15) of 2 µM PGE 2 . Incubation of cECs with PGE 2 lasted ~15 min. Data in B are means (column bars) ± SEM (error bars, ****p<0.0001, unpaired Student’s t-test). ( C ) Top: Schematic diagram showing the G q PCR-dependent hydrolysis of PIP 2 and the interventions used to test different components of the proposed pathway. Bottom: Summary data showing GSK101 (100 nM)-induced currents recorded at 100 mV in cECs dialyzed with 1 mM Mg-ATP. Currents were recorded in the absence and presence of 2 µM PGE 2 (orange shading), with or without (control) the indicated interventions. Concentrations (and application method): HC-067047, 1 µM (bath); diC8-PIP 2 , 10 µM (pipette), U73122, 10 µM (bath); U73343, 10 µM (bath); AH6809, 10 µM (bath); SC51322, 1 µM (bath); calphostin C, 0.5 µM (bath); Gö6976, 1 µM (bath); CPA, 30 µM (bath); BAPTA, 5.4 mM (pipette). For bath application, pharmacological agents were added 10–15 min before PGE 2 application. 10.7554/eLife.38689.025 Figure 5—source data 1. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.026 Figure 5—source data 2. Numerical data that were used to generate the chart in .

    Journal: eLife

    Article Title: PIP 2 depletion promotes TRPV4 channel activity in mouse brain capillary endothelial cells

    doi: 10.7554/eLife.38689

    Figure Lengend Snippet: ( A ) Representative current-voltage plots obtained from a cEC dialyzed with 1 mM Mg-ATP and treated consecutively with GSK101 (100 nM) and RuR (1 µM) followed by 2 µM PGE 2 . ( B ) Summary individual-value plot of GSK101-induced TRPV4 currents at 100 mV in cECs dialyzed with 1 mM Mg-ATP in the absence (black; n = 27) and presence (orange; n = 15) of 2 µM PGE 2 . Incubation of cECs with PGE 2 lasted ~15 min. Data in B are means (column bars) ± SEM (error bars, ****p<0.0001, unpaired Student’s t-test). ( C ) Top: Schematic diagram showing the G q PCR-dependent hydrolysis of PIP 2 and the interventions used to test different components of the proposed pathway. Bottom: Summary data showing GSK101 (100 nM)-induced currents recorded at 100 mV in cECs dialyzed with 1 mM Mg-ATP. Currents were recorded in the absence and presence of 2 µM PGE 2 (orange shading), with or without (control) the indicated interventions. Concentrations (and application method): HC-067047, 1 µM (bath); diC8-PIP 2 , 10 µM (pipette), U73122, 10 µM (bath); U73343, 10 µM (bath); AH6809, 10 µM (bath); SC51322, 1 µM (bath); calphostin C, 0.5 µM (bath); Gö6976, 1 µM (bath); CPA, 30 µM (bath); BAPTA, 5.4 mM (pipette). For bath application, pharmacological agents were added 10–15 min before PGE 2 application. 10.7554/eLife.38689.025 Figure 5—source data 1. Numerical data that were used to generate the chart in . 10.7554/eLife.38689.026 Figure 5—source data 2. Numerical data that were used to generate the chart in .

    Article Snippet: 1,2-Dioctanoyl phosphatidylinositol 4,5-bisphosphate sodium salt (diC8-PIP 2 ) and 1,2-dipalmitoyl phosphatidylinositol 4,5-bisphosphate sodium salt (diC16-PIP 2 ) were purchased from Cayman Chemical (USA).

    Techniques: Incubation, Transferring

    ( A ) Current-voltage relationship recorded in a cEC dialyzed with 1 mM Mg-ATP using voltage ramps (−100 to 100 mV). The circled numbers indicated the sequential and cumulative application of GSK101 (100 nM)+RuR (1 µM) followed by 2 µM PGE 2 and lastly, the TRPV4 blocker HC-067047 (1 µM). ( B ) Representative scatter plots of peak current amplitudes (at 100 mV) in three cECs dialyzed with 1 mM Mg-ATP and treated with 100 nM GSK101 followed by 2 µM PGE 2 (or PGE 2 first followed by GSK101). Note that PGE 2 -induced activation of TRPV4 current plateaued after ~13–15 min. ( C ) Scatter plots of normalized GSK101-induced outward currents (at 100 mV), normalized to the current at zero time, I 0 (when 2 µM PGE 2 was applied), in two cECs, one dialyzed with 1 mM Mg-ATP (orange symbols) and the other dialyzed with 1 mM Mg-ATP +10 µM diC8-PIP 2 (black symbols). Currents normalized to I 0 are presented before and after the application of PGE 2 for 35 min.

    Journal: eLife

    Article Title: PIP 2 depletion promotes TRPV4 channel activity in mouse brain capillary endothelial cells

    doi: 10.7554/eLife.38689

    Figure Lengend Snippet: ( A ) Current-voltage relationship recorded in a cEC dialyzed with 1 mM Mg-ATP using voltage ramps (−100 to 100 mV). The circled numbers indicated the sequential and cumulative application of GSK101 (100 nM)+RuR (1 µM) followed by 2 µM PGE 2 and lastly, the TRPV4 blocker HC-067047 (1 µM). ( B ) Representative scatter plots of peak current amplitudes (at 100 mV) in three cECs dialyzed with 1 mM Mg-ATP and treated with 100 nM GSK101 followed by 2 µM PGE 2 (or PGE 2 first followed by GSK101). Note that PGE 2 -induced activation of TRPV4 current plateaued after ~13–15 min. ( C ) Scatter plots of normalized GSK101-induced outward currents (at 100 mV), normalized to the current at zero time, I 0 (when 2 µM PGE 2 was applied), in two cECs, one dialyzed with 1 mM Mg-ATP (orange symbols) and the other dialyzed with 1 mM Mg-ATP +10 µM diC8-PIP 2 (black symbols). Currents normalized to I 0 are presented before and after the application of PGE 2 for 35 min.

    Article Snippet: 1,2-Dioctanoyl phosphatidylinositol 4,5-bisphosphate sodium salt (diC8-PIP 2 ) and 1,2-dipalmitoyl phosphatidylinositol 4,5-bisphosphate sodium salt (diC16-PIP 2 ) were purchased from Cayman Chemical (USA).

    Techniques: Activation Assay

    ( A ) Current-voltage relationship represent currents obtained from a cEC dialyzed with 0 µM diC8-PIP 2 and 0 mM Mg-ATP before (control) and after the application of 11,12-EET (1 µM) and RuR (1 µM), recorded using 300 ms voltage ramps (−100 to 100 mV). ( B ) Summary data showing 11,12-EET-induced outward currents at 100 mV in dialyzed cECs based on experiments in A (n = 6 cECs).

    Journal: eLife

    Article Title: PIP 2 depletion promotes TRPV4 channel activity in mouse brain capillary endothelial cells

    doi: 10.7554/eLife.38689

    Figure Lengend Snippet: ( A ) Current-voltage relationship represent currents obtained from a cEC dialyzed with 0 µM diC8-PIP 2 and 0 mM Mg-ATP before (control) and after the application of 11,12-EET (1 µM) and RuR (1 µM), recorded using 300 ms voltage ramps (−100 to 100 mV). ( B ) Summary data showing 11,12-EET-induced outward currents at 100 mV in dialyzed cECs based on experiments in A (n = 6 cECs).

    Article Snippet: 1,2-Dioctanoyl phosphatidylinositol 4,5-bisphosphate sodium salt (diC8-PIP 2 ) and 1,2-dipalmitoyl phosphatidylinositol 4,5-bisphosphate sodium salt (diC16-PIP 2 ) were purchased from Cayman Chemical (USA).

    Techniques: