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trpc1  (Alomone Labs)
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Fig. 1. Co-immunoprecipitation of TRPV4, <t>TRPC1</t> and KCa1.1(α) in the lysates of human LIMA VSMCs. A and B, TRPC1 with KCa1.1(α). C and D, TRPC1 with TRPV4. E and F, TRPV4 with KCa1.1(α). The data are representative of 3 to 4 experiments. The pulling antibody and the blotting antibody were indicated (+). Control immunoprecipitation (−) was performed using the preimmune IgG. IP, immunoprecipitation; Blot, immunoblot.
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Fig. 1. Co-immunoprecipitation of TRPV4, TRPC1 and KCa1.1(α) in the lysates of human LIMA VSMCs. A and B, TRPC1 with KCa1.1(α). C and D, TRPC1 with TRPV4. E and F, TRPV4 with KCa1.1(α). The data are representative of 3 to 4 experiments. The pulling antibody and the blotting antibody were indicated (+). Control immunoprecipitation (−) was performed using the preimmune IgG. IP, immunoprecipitation; Blot, immunoblot.

Journal: Biochimica et biophysica acta

Article Title: Epoxyeicosatrienoic acids act through TRPV4-TRPC1-KCa1.1 complex to induce smooth muscle membrane hyperpolarization and relaxation in human internal mammary arteries.

doi: 10.1016/j.bbadis.2014.12.010

Figure Lengend Snippet: Fig. 1. Co-immunoprecipitation of TRPV4, TRPC1 and KCa1.1(α) in the lysates of human LIMA VSMCs. A and B, TRPC1 with KCa1.1(α). C and D, TRPC1 with TRPV4. E and F, TRPV4 with KCa1.1(α). The data are representative of 3 to 4 experiments. The pulling antibody and the blotting antibody were indicated (+). Control immunoprecipitation (−) was performed using the preimmune IgG. IP, immunoprecipitation; Blot, immunoblot.

Article Snippet: The primary rabbit antibodies against KCa1.1(α) (APC-021), TRPC1 (ACC-010), and TRPV4 (ACC-034) were from Alomone Labs. T1E3 was raised by us from rabbits.

Techniques: Immunoprecipitation, Control, Western Blot

Fig. 5. Membrane potential changes in HEK293 cells that over-expressed with TRPV4, TRPC1 and KCa1.1. A and B, representative traces illustrating the hyperpolarizing responses to 300 nmol/L 11,12-EET (A), 5 μmol/L 4a-PDD (B) and 100 nmol/L GSK1016790A (C). D–F, summary of data showing the maximal hyperpolarization to 300 nmol/L 11,12-EET (D), 5 μmol/L 4a- PDD (E) and 100 nmol/L GSK1016790A (F) under different conditions. C1mut-pore, TRPC1 were replaced by a TRPC1 pore-dead mutant C1mut-pore. Values are means ± SE (n = 5 to 9). **, P b 0.01 compared to the controls (non-transfected); ##, P b 0.01 compared to the transfected cells without inhibitors; $$, P b 0.01 compared to preimmune IgG.

Journal: Biochimica et biophysica acta

Article Title: Epoxyeicosatrienoic acids act through TRPV4-TRPC1-KCa1.1 complex to induce smooth muscle membrane hyperpolarization and relaxation in human internal mammary arteries.

doi: 10.1016/j.bbadis.2014.12.010

Figure Lengend Snippet: Fig. 5. Membrane potential changes in HEK293 cells that over-expressed with TRPV4, TRPC1 and KCa1.1. A and B, representative traces illustrating the hyperpolarizing responses to 300 nmol/L 11,12-EET (A), 5 μmol/L 4a-PDD (B) and 100 nmol/L GSK1016790A (C). D–F, summary of data showing the maximal hyperpolarization to 300 nmol/L 11,12-EET (D), 5 μmol/L 4a- PDD (E) and 100 nmol/L GSK1016790A (F) under different conditions. C1mut-pore, TRPC1 were replaced by a TRPC1 pore-dead mutant C1mut-pore. Values are means ± SE (n = 5 to 9). **, P b 0.01 compared to the controls (non-transfected); ##, P b 0.01 compared to the transfected cells without inhibitors; $$, P b 0.01 compared to preimmune IgG.

Article Snippet: The primary rabbit antibodies against KCa1.1(α) (APC-021), TRPC1 (ACC-010), and TRPV4 (ACC-034) were from Alomone Labs. T1E3 was raised by us from rabbits.

Techniques: Membrane, Mutagenesis, Transfection

Fig. 4. Requirement of TRPC1 for TRPV4 and KCa1.1 interaction. A–D, co-immunoprecipitation of TRPV4 and KCa1.1(α) in the lysates of HEK293 cells. A and B, HEK293 cells that over- expressed with TRPV4, TRPC1 and KCa1.1. C and D, HEK293 cells that over-expressed with TRPV4 and KCa1.1 but without TRPC1. The right lanes in C and D were immunoblots using cell lysates without immunoprecipitation. The pulling antibody and the blotting antibody were indicated (+). Control immunoprecipitation (−) was performed using the preimmune IgG. IP, immunoprecipitation; Blot, immunoblot. n = 3 to 4 experiments. E–F, the maximal hyperpolarization in response to 300 nmol/L 11,12-EET (E) and 5 μmol/L 4α-PDD (F) in HEK293 cells that were transfected with indicated constructs. C1 stands for TRPC1. V4 stands for TRPV4. Values are means ± SE (n = 5 to 9). **P b 0.01 compared to the controls (non-transfected), ##P b 0.01 compared to C1 + V4 + KCa1.1.

Journal: Biochimica et biophysica acta

Article Title: Epoxyeicosatrienoic acids act through TRPV4-TRPC1-KCa1.1 complex to induce smooth muscle membrane hyperpolarization and relaxation in human internal mammary arteries.

doi: 10.1016/j.bbadis.2014.12.010

Figure Lengend Snippet: Fig. 4. Requirement of TRPC1 for TRPV4 and KCa1.1 interaction. A–D, co-immunoprecipitation of TRPV4 and KCa1.1(α) in the lysates of HEK293 cells. A and B, HEK293 cells that over- expressed with TRPV4, TRPC1 and KCa1.1. C and D, HEK293 cells that over-expressed with TRPV4 and KCa1.1 but without TRPC1. The right lanes in C and D were immunoblots using cell lysates without immunoprecipitation. The pulling antibody and the blotting antibody were indicated (+). Control immunoprecipitation (−) was performed using the preimmune IgG. IP, immunoprecipitation; Blot, immunoblot. n = 3 to 4 experiments. E–F, the maximal hyperpolarization in response to 300 nmol/L 11,12-EET (E) and 5 μmol/L 4α-PDD (F) in HEK293 cells that were transfected with indicated constructs. C1 stands for TRPC1. V4 stands for TRPV4. Values are means ± SE (n = 5 to 9). **P b 0.01 compared to the controls (non-transfected), ##P b 0.01 compared to C1 + V4 + KCa1.1.

Article Snippet: The primary rabbit antibodies against KCa1.1(α) (APC-021), TRPC1 (ACC-010), and TRPV4 (ACC-034) were from Alomone Labs. T1E3 was raised by us from rabbits.

Techniques: Immunoprecipitation, Western Blot, Control, Transfection, Construct