Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Expression of TagBFP2 adenoviral constructs in NRVMs. Fluorescent imaging of TagBFP2 in cells expressing (A) TRPC1-TagBFP2, (B) TagBFP2, (C) scrambled shRNA-TagBFP2, and (D) shRNA-TRPC1-TagBFP2. Scale bars = 20 µm.

Article Snippet: Mean fold change of TRPC1 expression compared to WT NRVMs is significantly greater in TRPC1-TagBFP2 cells than TagBFP2, scrambled shRNA-TagBFP2, and shRNA-TRPC1-TagBFP2 cells for the primer targeting exon 3-4 (Rn00677552_m1, all p < 0.001), the primer targeting exon 6-7 (Rn00677554_m1, all p < 0.001), the primer targeting exon 8-9 (Rn00585625_m1, all p ≤ 0.001) and the primer targeting exon 10-11 (Rn00677549_g1, all p < 0.001).

Techniques: Expressing, Construct, Imaging, shRNA

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Mean fold change of TRPC1 mRNA from WT NRVMs of all RT-qPCR primers.

Article Snippet: Mean fold change of TRPC1 expression compared to WT NRVMs is significantly greater in TRPC1-TagBFP2 cells than TagBFP2, scrambled shRNA-TagBFP2, and shRNA-TRPC1-TagBFP2 cells for the primer targeting exon 3-4 (Rn00677552_m1, all p < 0.001), the primer targeting exon 6-7 (Rn00677554_m1, all p < 0.001), the primer targeting exon 8-9 (Rn00585625_m1, all p ≤ 0.001) and the primer targeting exon 10-11 (Rn00677549_g1, all p < 0.001).

Techniques: shRNA

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Mean fold change of TRPC1 mRNA expression of infected NRVMs from WT NRVMs quantified by RT-qPCR with primers targeting rat TRPC1 (A) exon 3-4 (Rn00677552_m1) (B) exon 6-7 (Rn00677554_m1) (C) exon 8-9 (Rn00585625_m1) (D) exon 10-11 (Rn00677549_g1) and (E) exon 11-12 (Rn01447000_m1) ( n = 3 litters). Brackets mark significant differences ( p < 0.05).

Article Snippet: Mean fold change of TRPC1 expression compared to WT NRVMs is significantly greater in TRPC1-TagBFP2 cells than TagBFP2, scrambled shRNA-TagBFP2, and shRNA-TRPC1-TagBFP2 cells for the primer targeting exon 3-4 (Rn00677552_m1, all p < 0.001), the primer targeting exon 6-7 (Rn00677554_m1, all p < 0.001), the primer targeting exon 8-9 (Rn00585625_m1, all p ≤ 0.001) and the primer targeting exon 10-11 (Rn00677549_g1, all p < 0.001).

Techniques: Expressing, Infection, Quantitative RT-PCR

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Super-resolution images of NRVMs infected with TRPC1-TagBFP2 (A–F) and TagBFP2 (G – L) . TRPC1-TagBFP2 cells exhibit a striated arrangement of both (A) TagBFP2 and (B) SERCA2 fluorescence. (C) Colocalization of TagBFP2 and SERCA2 in TRPC1-TagBFP2 cells is high, shown in white. (D–F) Zoomed in region from (A–C) depicted by white box in (C) . (G) TagBFP2 fluorescence in control cells is distributed throughout the cell, with more dense concentration near sarcomere structures. (H) SERCA2 fluorescence in TagBFP2 control cells follows a network, with (I) minimal colocalization with TagBFP2. (J–L) Zoomed in region from (G–I) depicted by white box in (I) . Scale bars in (C,I) = 10 µm. Scale bars in (F,L) = 3 µm.

Article Snippet: Mean fold change of TRPC1 expression compared to WT NRVMs is significantly greater in TRPC1-TagBFP2 cells than TagBFP2, scrambled shRNA-TagBFP2, and shRNA-TRPC1-TagBFP2 cells for the primer targeting exon 3-4 (Rn00677552_m1, all p < 0.001), the primer targeting exon 6-7 (Rn00677554_m1, all p < 0.001), the primer targeting exon 8-9 (Rn00585625_m1, all p ≤ 0.001) and the primer targeting exon 10-11 (Rn00677549_g1, all p < 0.001).

Techniques: Infection, Fluorescence, Control, Concentration Assay

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Quantification of colocalization from super-resolution images of NRVMs infected with TRPC1-TagBFP2 or TagBFP2. (A) Average distribution of nearest neighbor distances (NND) of SERCA2 from TagBFP2 in TRPC1-TagBFP2 cells and TagBFP2 control cells. (B) Mean NND in TRPC1-TagBFP2 cells (98.08 ± 4.98 nm) is significantly lower than in TagBFP2 control cells (145.93 ± 5.55 nm) ( n = 8 cells). Bracket marks significant difference ( p < 0.001).

Article Snippet: Mean fold change of TRPC1 expression compared to WT NRVMs is significantly greater in TRPC1-TagBFP2 cells than TagBFP2, scrambled shRNA-TagBFP2, and shRNA-TRPC1-TagBFP2 cells for the primer targeting exon 3-4 (Rn00677552_m1, all p < 0.001), the primer targeting exon 6-7 (Rn00677554_m1, all p < 0.001), the primer targeting exon 8-9 (Rn00585625_m1, all p ≤ 0.001) and the primer targeting exon 10-11 (Rn00677549_g1, all p < 0.001).

Techniques: Infection, Control

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Super-resolution images of GCEPIAer-SNAP construct in TRPC1-TagBFP2 cells. (A) SNAP-Cell 647-SiR signal exhibits a network pattern strikingly similar to the (B) tRFP signal corresponding to TRPC1-TagBFP2. (C) The patterns exhibited from markers for both the GCEPIAer-SNAP construct and TRPC1-TagBFP2 construct reveal abundant overlap. Scale bar = 5 µm.

Article Snippet: Mean fold change of TRPC1 expression compared to WT NRVMs is significantly greater in TRPC1-TagBFP2 cells than TagBFP2, scrambled shRNA-TagBFP2, and shRNA-TRPC1-TagBFP2 cells for the primer targeting exon 3-4 (Rn00677552_m1, all p < 0.001), the primer targeting exon 6-7 (Rn00677554_m1, all p < 0.001), the primer targeting exon 8-9 (Rn00585625_m1, all p ≤ 0.001) and the primer targeting exon 10-11 (Rn00677549_g1, all p < 0.001).

Techniques: Construct

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Calibrated [Ca 2+ ] SR from fluorescent calcium imaging of NRVMs. Representative ratiometric images of the GCEPIAer signal over the SNAP signal in NRVMs infected with (A) TRPC1-TagBFP2 under 0% strain, (B) TagBFP2 under 0% strain, (C) shRNA-TRPC1-TagBFP2 under 0% strain, (D) TRPC1-TagBFP2 under 10% strain, (E) TagBFP2 under 10% strain, and (F) shRNA-TRPC1-TagBFP2 under 10% strain. Scale bars = 20 µm. (G) Example [Ca 2+ ] SR transients from TRPC1-TagBFP2, TagBFP2 and shRNA-TRPC1-TagBFP2 infected un stretched and stretched NRVMs. (H) Scatter plot of diastolic [Ca 2+ ] SR of each infection group with 0 or 10% stretch averaged by membrane; points correspond to the average value of all cells on each membrane and bars represent mean and SEM determined by multilevel mixed-effects model. Diastolic [Ca 2+ ] SR in unstretched NRVMs is significantly lower in TRPC1-TagBFP2 and TagBFP2 cells compared to shRNA-TRPC1-TagBFP2 cells. Sustained 10% stretch significantly lowered [Ca 2+ ] SR in TRPC1-TagBFP2 infected NRVMs. Brackets mark significant differences ( p < 0.05).

Article Snippet: Mean fold change of TRPC1 expression compared to WT NRVMs is significantly greater in TRPC1-TagBFP2 cells than TagBFP2, scrambled shRNA-TagBFP2, and shRNA-TRPC1-TagBFP2 cells for the primer targeting exon 3-4 (Rn00677552_m1, all p < 0.001), the primer targeting exon 6-7 (Rn00677554_m1, all p < 0.001), the primer targeting exon 8-9 (Rn00585625_m1, all p ≤ 0.001) and the primer targeting exon 10-11 (Rn00677549_g1, all p < 0.001).

Techniques: Imaging, Infection, shRNA, Membrane

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Data extracted from calibrated [Ca 2+ ]SR transients of NRVMs.

Article Snippet: Mean fold change of TRPC1 expression compared to WT NRVMs is significantly greater in TRPC1-TagBFP2 cells than TagBFP2, scrambled shRNA-TagBFP2, and shRNA-TRPC1-TagBFP2 cells for the primer targeting exon 3-4 (Rn00677552_m1, all p < 0.001), the primer targeting exon 6-7 (Rn00677554_m1, all p < 0.001), the primer targeting exon 8-9 (Rn00585625_m1, all p ≤ 0.001) and the primer targeting exon 10-11 (Rn00677549_g1, all p < 0.001).

Techniques: shRNA

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Expression of TagBFP2 adenoviral constructs in NRVMs. Fluorescent imaging of TagBFP2 in cells expressing (A) TRPC1-TagBFP2, (B) TagBFP2, (C) scrambled shRNA-TagBFP2, and (D) shRNA-TRPC1-TagBFP2. Scale bars = 20 µm.

Article Snippet: Rn00677549_g1 , Exon 10–11 , 2,427.12 ± 261.53 , 0.91 ± 0.08 , 0.97 ± 0.19 , 0.26 ± 0.07.

Techniques: Expressing, Construct, Imaging, shRNA

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Mean fold change of TRPC1 mRNA from WT NRVMs of all RT-qPCR primers.

Article Snippet: Rn00677549_g1 , Exon 10–11 , 2,427.12 ± 261.53 , 0.91 ± 0.08 , 0.97 ± 0.19 , 0.26 ± 0.07.

Techniques: shRNA

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Mean fold change of TRPC1 mRNA expression of infected NRVMs from WT NRVMs quantified by RT-qPCR with primers targeting rat TRPC1 (A) exon 3-4 (Rn00677552_m1) (B) exon 6-7 (Rn00677554_m1) (C) exon 8-9 (Rn00585625_m1) (D) exon 10-11 (Rn00677549_g1) and (E) exon 11-12 (Rn01447000_m1) ( n = 3 litters). Brackets mark significant differences ( p < 0.05).

Article Snippet: Rn00677549_g1 , Exon 10–11 , 2,427.12 ± 261.53 , 0.91 ± 0.08 , 0.97 ± 0.19 , 0.26 ± 0.07.

Techniques: Expressing, Infection, Quantitative RT-PCR

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Super-resolution images of NRVMs infected with TRPC1-TagBFP2 (A–F) and TagBFP2 (G – L) . TRPC1-TagBFP2 cells exhibit a striated arrangement of both (A) TagBFP2 and (B) SERCA2 fluorescence. (C) Colocalization of TagBFP2 and SERCA2 in TRPC1-TagBFP2 cells is high, shown in white. (D–F) Zoomed in region from (A–C) depicted by white box in (C) . (G) TagBFP2 fluorescence in control cells is distributed throughout the cell, with more dense concentration near sarcomere structures. (H) SERCA2 fluorescence in TagBFP2 control cells follows a network, with (I) minimal colocalization with TagBFP2. (J–L) Zoomed in region from (G–I) depicted by white box in (I) . Scale bars in (C,I) = 10 µm. Scale bars in (F,L) = 3 µm.

Article Snippet: Rn00677549_g1 , Exon 10–11 , 2,427.12 ± 261.53 , 0.91 ± 0.08 , 0.97 ± 0.19 , 0.26 ± 0.07.

Techniques: Infection, Fluorescence, Control, Concentration Assay

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Quantification of colocalization from super-resolution images of NRVMs infected with TRPC1-TagBFP2 or TagBFP2. (A) Average distribution of nearest neighbor distances (NND) of SERCA2 from TagBFP2 in TRPC1-TagBFP2 cells and TagBFP2 control cells. (B) Mean NND in TRPC1-TagBFP2 cells (98.08 ± 4.98 nm) is significantly lower than in TagBFP2 control cells (145.93 ± 5.55 nm) ( n = 8 cells). Bracket marks significant difference ( p < 0.001).

Article Snippet: Rn00677549_g1 , Exon 10–11 , 2,427.12 ± 261.53 , 0.91 ± 0.08 , 0.97 ± 0.19 , 0.26 ± 0.07.

Techniques: Infection, Control

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Super-resolution images of GCEPIAer-SNAP construct in TRPC1-TagBFP2 cells. (A) SNAP-Cell 647-SiR signal exhibits a network pattern strikingly similar to the (B) tRFP signal corresponding to TRPC1-TagBFP2. (C) The patterns exhibited from markers for both the GCEPIAer-SNAP construct and TRPC1-TagBFP2 construct reveal abundant overlap. Scale bar = 5 µm.

Article Snippet: Rn00677549_g1 , Exon 10–11 , 2,427.12 ± 261.53 , 0.91 ± 0.08 , 0.97 ± 0.19 , 0.26 ± 0.07.

Techniques: Construct

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Calibrated [Ca 2+ ] SR from fluorescent calcium imaging of NRVMs. Representative ratiometric images of the GCEPIAer signal over the SNAP signal in NRVMs infected with (A) TRPC1-TagBFP2 under 0% strain, (B) TagBFP2 under 0% strain, (C) shRNA-TRPC1-TagBFP2 under 0% strain, (D) TRPC1-TagBFP2 under 10% strain, (E) TagBFP2 under 10% strain, and (F) shRNA-TRPC1-TagBFP2 under 10% strain. Scale bars = 20 µm. (G) Example [Ca 2+ ] SR transients from TRPC1-TagBFP2, TagBFP2 and shRNA-TRPC1-TagBFP2 infected un stretched and stretched NRVMs. (H) Scatter plot of diastolic [Ca 2+ ] SR of each infection group with 0 or 10% stretch averaged by membrane; points correspond to the average value of all cells on each membrane and bars represent mean and SEM determined by multilevel mixed-effects model. Diastolic [Ca 2+ ] SR in unstretched NRVMs is significantly lower in TRPC1-TagBFP2 and TagBFP2 cells compared to shRNA-TRPC1-TagBFP2 cells. Sustained 10% stretch significantly lowered [Ca 2+ ] SR in TRPC1-TagBFP2 infected NRVMs. Brackets mark significant differences ( p < 0.05).

Article Snippet: Rn00677549_g1 , Exon 10–11 , 2,427.12 ± 261.53 , 0.91 ± 0.08 , 0.97 ± 0.19 , 0.26 ± 0.07.

Techniques: Imaging, Infection, shRNA, Membrane

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Data extracted from calibrated [Ca 2+ ]SR transients of NRVMs.

Article Snippet: Rn00677549_g1 , Exon 10–11 , 2,427.12 ± 261.53 , 0.91 ± 0.08 , 0.97 ± 0.19 , 0.26 ± 0.07.

Techniques: shRNA

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Expression of TagBFP2 adenoviral constructs in NRVMs. Fluorescent imaging of TagBFP2 in cells expressing (A) TRPC1-TagBFP2, (B) TagBFP2, (C) scrambled shRNA-TagBFP2, and (D) shRNA-TRPC1-TagBFP2. Scale bars = 20 µm.

Article Snippet: We applied five TaqManTM primers, with FAM-MGB dye, for TRPC1 ( Rn00677552_m1 , Rn00677554_m1 , Rn00585625_m1 , Rn00677549_g1 , Rn01447000_m1 , Thermo Fisher Scientific) and used the TaqMan ® Fast Advanced Master Mix (Thermo Fisher Scientific) with a total volume of 10 μl.

Techniques: Expressing, Construct, Imaging, shRNA

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Mean fold change of TRPC1 mRNA from WT NRVMs of all RT-qPCR primers.

Article Snippet: We applied five TaqManTM primers, with FAM-MGB dye, for TRPC1 ( Rn00677552_m1 , Rn00677554_m1 , Rn00585625_m1 , Rn00677549_g1 , Rn01447000_m1 , Thermo Fisher Scientific) and used the TaqMan ® Fast Advanced Master Mix (Thermo Fisher Scientific) with a total volume of 10 μl.

Techniques: shRNA

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Mean fold change of TRPC1 mRNA expression of infected NRVMs from WT NRVMs quantified by RT-qPCR with primers targeting rat TRPC1 (A) exon 3-4 (Rn00677552_m1) (B) exon 6-7 (Rn00677554_m1) (C) exon 8-9 (Rn00585625_m1) (D) exon 10-11 (Rn00677549_g1) and (E) exon 11-12 (Rn01447000_m1) ( n = 3 litters). Brackets mark significant differences ( p < 0.05).

Article Snippet: We applied five TaqManTM primers, with FAM-MGB dye, for TRPC1 ( Rn00677552_m1 , Rn00677554_m1 , Rn00585625_m1 , Rn00677549_g1 , Rn01447000_m1 , Thermo Fisher Scientific) and used the TaqMan ® Fast Advanced Master Mix (Thermo Fisher Scientific) with a total volume of 10 μl.

Techniques: Expressing, Infection, Quantitative RT-PCR

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Super-resolution images of NRVMs infected with TRPC1-TagBFP2 (A–F) and TagBFP2 (G – L) . TRPC1-TagBFP2 cells exhibit a striated arrangement of both (A) TagBFP2 and (B) SERCA2 fluorescence. (C) Colocalization of TagBFP2 and SERCA2 in TRPC1-TagBFP2 cells is high, shown in white. (D–F) Zoomed in region from (A–C) depicted by white box in (C) . (G) TagBFP2 fluorescence in control cells is distributed throughout the cell, with more dense concentration near sarcomere structures. (H) SERCA2 fluorescence in TagBFP2 control cells follows a network, with (I) minimal colocalization with TagBFP2. (J–L) Zoomed in region from (G–I) depicted by white box in (I) . Scale bars in (C,I) = 10 µm. Scale bars in (F,L) = 3 µm.

Article Snippet: We applied five TaqManTM primers, with FAM-MGB dye, for TRPC1 ( Rn00677552_m1 , Rn00677554_m1 , Rn00585625_m1 , Rn00677549_g1 , Rn01447000_m1 , Thermo Fisher Scientific) and used the TaqMan ® Fast Advanced Master Mix (Thermo Fisher Scientific) with a total volume of 10 μl.

Techniques: Infection, Fluorescence, Control, Concentration Assay

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Quantification of colocalization from super-resolution images of NRVMs infected with TRPC1-TagBFP2 or TagBFP2. (A) Average distribution of nearest neighbor distances (NND) of SERCA2 from TagBFP2 in TRPC1-TagBFP2 cells and TagBFP2 control cells. (B) Mean NND in TRPC1-TagBFP2 cells (98.08 ± 4.98 nm) is significantly lower than in TagBFP2 control cells (145.93 ± 5.55 nm) ( n = 8 cells). Bracket marks significant difference ( p < 0.001).

Article Snippet: We applied five TaqManTM primers, with FAM-MGB dye, for TRPC1 ( Rn00677552_m1 , Rn00677554_m1 , Rn00585625_m1 , Rn00677549_g1 , Rn01447000_m1 , Thermo Fisher Scientific) and used the TaqMan ® Fast Advanced Master Mix (Thermo Fisher Scientific) with a total volume of 10 μl.

Techniques: Infection, Control

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Super-resolution images of GCEPIAer-SNAP construct in TRPC1-TagBFP2 cells. (A) SNAP-Cell 647-SiR signal exhibits a network pattern strikingly similar to the (B) tRFP signal corresponding to TRPC1-TagBFP2. (C) The patterns exhibited from markers for both the GCEPIAer-SNAP construct and TRPC1-TagBFP2 construct reveal abundant overlap. Scale bar = 5 µm.

Article Snippet: We applied five TaqManTM primers, with FAM-MGB dye, for TRPC1 ( Rn00677552_m1 , Rn00677554_m1 , Rn00585625_m1 , Rn00677549_g1 , Rn01447000_m1 , Thermo Fisher Scientific) and used the TaqMan ® Fast Advanced Master Mix (Thermo Fisher Scientific) with a total volume of 10 μl.

Techniques: Construct

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Calibrated [Ca 2+ ] SR from fluorescent calcium imaging of NRVMs. Representative ratiometric images of the GCEPIAer signal over the SNAP signal in NRVMs infected with (A) TRPC1-TagBFP2 under 0% strain, (B) TagBFP2 under 0% strain, (C) shRNA-TRPC1-TagBFP2 under 0% strain, (D) TRPC1-TagBFP2 under 10% strain, (E) TagBFP2 under 10% strain, and (F) shRNA-TRPC1-TagBFP2 under 10% strain. Scale bars = 20 µm. (G) Example [Ca 2+ ] SR transients from TRPC1-TagBFP2, TagBFP2 and shRNA-TRPC1-TagBFP2 infected un stretched and stretched NRVMs. (H) Scatter plot of diastolic [Ca 2+ ] SR of each infection group with 0 or 10% stretch averaged by membrane; points correspond to the average value of all cells on each membrane and bars represent mean and SEM determined by multilevel mixed-effects model. Diastolic [Ca 2+ ] SR in unstretched NRVMs is significantly lower in TRPC1-TagBFP2 and TagBFP2 cells compared to shRNA-TRPC1-TagBFP2 cells. Sustained 10% stretch significantly lowered [Ca 2+ ] SR in TRPC1-TagBFP2 infected NRVMs. Brackets mark significant differences ( p < 0.05).

Article Snippet: We applied five TaqManTM primers, with FAM-MGB dye, for TRPC1 ( Rn00677552_m1 , Rn00677554_m1 , Rn00585625_m1 , Rn00677549_g1 , Rn01447000_m1 , Thermo Fisher Scientific) and used the TaqMan ® Fast Advanced Master Mix (Thermo Fisher Scientific) with a total volume of 10 μl.

Techniques: Imaging, Infection, shRNA, Membrane

Journal: Frontiers in Physiology

Article Title: TRPC1 channels underlie stretch-modulated sarcoplasmic reticulum calcium leak in cardiomyocytes

doi: 10.3389/fphys.2022.1056657

Figure Lengend Snippet: Data extracted from calibrated [Ca 2+ ]SR transients of NRVMs.

Article Snippet: We applied five TaqManTM primers, with FAM-MGB dye, for TRPC1 ( Rn00677552_m1 , Rn00677554_m1 , Rn00585625_m1 , Rn00677549_g1 , Rn01447000_m1 , Thermo Fisher Scientific) and used the TaqMan ® Fast Advanced Master Mix (Thermo Fisher Scientific) with a total volume of 10 μl.

Techniques: shRNA

Journal: Marine Drugs

Article Title: Selective Inhibition of Liver Cancer Cells Using Venom Peptide

doi: 10.3390/md17100587

Figure Lengend Snippet: Potential mechanism of Tv1 antitumor activity inhibits COX2 and PGE2 function. In our model of Tv1 antitumor activity in liver cancer cells, overexpression of TRP channels (TRPC6 and V6) stimulates COX-2-dependent PGE2 production via enhanced [Ca 2+ ] dynamics. Influx of Ca 2+ can occur through voltage gated (VGC), receptor operated (ROC), and store operated (SOC) calcium channels. Transient receptor potential (TRP) channels contribute to store operated calcium (SOC) channels. Ca 2+ -dependent transcription factor NFAT is activated via dephosphorylation by calcineurin, which is activated upon binding of Ca 2+ /calmodulin. Ubiquitously present transcription factor NFAT regulates COX-2 expression and further prostaglandin E2 (PGE2) release in different cancer cells and its activation occurs through Ca 2+ influx associated with TRPC1-, TRPC3-, or TRPC6-associated SOC or ROC activities. PGE2 release plays multiple roles in cancer as shown. Upon Tv1 application to liver cancer (HCC) cells the pictured downstream pathways leading to proliferation inhibition and apoptosis are encountered.

Article Snippet: Rabbit polyclonal Anti-HERG (Kv11.1)-APC109, Anti TRPV6-ACC036, Anti TRPV2-ACC039, Anti TRPC1-ACC-118 were purchased from Alomone lab, Israel.

Techniques: Activity Assay, Over Expression, De-Phosphorylation Assay, Binding Assay, Expressing, Activation Assay, Inhibition

Journal: Scientific Reports

Article Title: Differential regulation of TRP channel gene and protein expression by intervertebral disc degeneration and back pain

doi: 10.1038/s41598-019-55212-9

Figure Lengend Snippet: TaqMan primers used for the qPCR analysis.

Article Snippet: TRPC1 , Transient receptor potential channel subfamily C member 1 , Hs00608195_m1.

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Trpc6 Promotes Doxorubicin-Induced Cardiomyopathy in Male Mice With Pleiotropic Differences Between Males and Females

doi: 10.3389/fcvm.2021.757784

Figure Lengend Snippet: Other Trpc gene expression is not altered in Trpc6 -deficient males after doxorubicin treatment. Male wild-type (WT) and Trpc6 -deficient (KO) mice were treated with 100 mL saline (CON) or 4 mg/kg doxorubicin (DOX) on days 1, 3, 5, 7, 9, 11 for a cumulative dose of 24 mg/kg. Cardiac gene expression shown as a fold change (FC) relative to WT control for (A) Trpc6 , (B) Trpc1 , (C) Trpc3 or (D) Rcan1 at day 21. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. (A) Data shown as mean ± SEM using 2 tailed Student's t test with 10 mice/ group. (B–D) Data shown as mean ± SEM using one-way ANOVA with Tukey's multiple comparisons test with 4–10 mice/ group.

Article Snippet: The following primer/probe sets were purchased from Applied Biosystems: Trpc1 (Mm00441975_m1), Trpc3 (Mm00444690_m1), Trpc6 (Mm01176083_m1), Myh7 (Mm00600555_m1), Tnni3 (Mm00437164_m1) and Rcan1 (Mm01213406_m1).

Techniques: Gene Expression, Saline, Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: Trpc6 Promotes Doxorubicin-Induced Cardiomyopathy in Male Mice With Pleiotropic Differences Between Males and Females

doi: 10.3389/fcvm.2021.757784

Figure Lengend Snippet: Trpc6 expression is reduced by doxorubicin in female wild-type mice while Trpc1, Trpc3 and Rcan1 expression is reduced by Trpc6 deficiency. Female wild-type (WT) and Trpc6 -deficient (KO) mice were treated with 100 mL saline (CON) or 4 mg/kg doxorubicin (DOX) on days 1, 3, 5, 7, 9, 11 for a cumulative dose of 24 mg/kg. Cardiac gene expression shown as a fold change (FC) relative to WT control (A) Trpc6 , (B) Trpc1 , (C) Trpc3 and (D) Rcan1 at day 21. * p < 0.05, ** p < 0.01, **** p < 0.0001. Data shown as mean ± SEM using one-way ANOVA with Tukey's multiple comparisons with 7–12 mice/ group.

Article Snippet: The following primer/probe sets were purchased from Applied Biosystems: Trpc1 (Mm00441975_m1), Trpc3 (Mm00444690_m1), Trpc6 (Mm01176083_m1), Myh7 (Mm00600555_m1), Tnni3 (Mm00437164_m1) and Rcan1 (Mm01213406_m1).

Techniques: Expressing, Saline, Gene Expression, Control