Structured Review

Abcam anti p ryr2
MiR-17-3p was the downstream target of circ-HIPK3. (A) Results of WB showing that <t>p-RyR2</t> and p-PLN increased in circ-HIPK3 overexpressed NMCMs and vice versa. (B) QRT-PCR showing that PLN and <t>RyR2</t> changed little in circ-HIPK3 over- or under- expressed NMCMs. NS = not significant. (C) Schematic image showing that seven possible binding sites of miR-17-3p to circ-HIPK3 were found. (D) Dual-luciferase reporter gene assay showed miR-17-3p could decrease the fluorescence density of NMCMs transfected with pmirGLO- wt -circ-HIPK3. * p<0.05 . (E) FISH assay showed circ-HIPK3 (red) can interact with miR-17-3p (green), which mainly were around the nucleus (blue). (F) The level of p-RyR2 and p-PLN were downregulated by miR-17-3p mimic or upregulated by inhibitor. (G) The peak FI of fluo-3 decreased in miR-17-3p transfected NMCMs and increased in inhibitor transfected NMCMs. ** p < 0.01, n = 15. (H) The FDHM of NMCMs activated by carbamylcholine was not affected by miR-17-3p or inhibitor. NS = not significant, n = 15. (I) The miR-17-3p can decrease the FI of 530nm and increase the 475nm in NMCMs; Inhibitor can increase the FI of 530 and decrease the 475nm. * p < 0.05, n = 15. (J) Both miR-17-3p and inhibitor had no effects on PLN and RyR2 in genetic level. NS = not significant.
Anti P Ryr2, supplied by Abcam, 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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anti p ryr2 - by Bioz Stars, 2023-01
86/100 stars

Images

1) Product Images from "Circ-HIPK3 Strengthens the Effects of Adrenaline in Heart Failure by MiR-17-3p - ADCY6 Axis"

Article Title: Circ-HIPK3 Strengthens the Effects of Adrenaline in Heart Failure by MiR-17-3p - ADCY6 Axis

Journal: International Journal of Biological Sciences

doi: 10.7150/ijbs.36149

MiR-17-3p was the downstream target of circ-HIPK3. (A) Results of WB showing that p-RyR2 and p-PLN increased in circ-HIPK3 overexpressed NMCMs and vice versa. (B) QRT-PCR showing that PLN and RyR2 changed little in circ-HIPK3 over- or under- expressed NMCMs. NS = not significant. (C) Schematic image showing that seven possible binding sites of miR-17-3p to circ-HIPK3 were found. (D) Dual-luciferase reporter gene assay showed miR-17-3p could decrease the fluorescence density of NMCMs transfected with pmirGLO- wt -circ-HIPK3. * p<0.05 . (E) FISH assay showed circ-HIPK3 (red) can interact with miR-17-3p (green), which mainly were around the nucleus (blue). (F) The level of p-RyR2 and p-PLN were downregulated by miR-17-3p mimic or upregulated by inhibitor. (G) The peak FI of fluo-3 decreased in miR-17-3p transfected NMCMs and increased in inhibitor transfected NMCMs. ** p < 0.01, n = 15. (H) The FDHM of NMCMs activated by carbamylcholine was not affected by miR-17-3p or inhibitor. NS = not significant, n = 15. (I) The miR-17-3p can decrease the FI of 530nm and increase the 475nm in NMCMs; Inhibitor can increase the FI of 530 and decrease the 475nm. * p < 0.05, n = 15. (J) Both miR-17-3p and inhibitor had no effects on PLN and RyR2 in genetic level. NS = not significant.
Figure Legend Snippet: MiR-17-3p was the downstream target of circ-HIPK3. (A) Results of WB showing that p-RyR2 and p-PLN increased in circ-HIPK3 overexpressed NMCMs and vice versa. (B) QRT-PCR showing that PLN and RyR2 changed little in circ-HIPK3 over- or under- expressed NMCMs. NS = not significant. (C) Schematic image showing that seven possible binding sites of miR-17-3p to circ-HIPK3 were found. (D) Dual-luciferase reporter gene assay showed miR-17-3p could decrease the fluorescence density of NMCMs transfected with pmirGLO- wt -circ-HIPK3. * p<0.05 . (E) FISH assay showed circ-HIPK3 (red) can interact with miR-17-3p (green), which mainly were around the nucleus (blue). (F) The level of p-RyR2 and p-PLN were downregulated by miR-17-3p mimic or upregulated by inhibitor. (G) The peak FI of fluo-3 decreased in miR-17-3p transfected NMCMs and increased in inhibitor transfected NMCMs. ** p < 0.01, n = 15. (H) The FDHM of NMCMs activated by carbamylcholine was not affected by miR-17-3p or inhibitor. NS = not significant, n = 15. (I) The miR-17-3p can decrease the FI of 530nm and increase the 475nm in NMCMs; Inhibitor can increase the FI of 530 and decrease the 475nm. * p < 0.05, n = 15. (J) Both miR-17-3p and inhibitor had no effects on PLN and RyR2 in genetic level. NS = not significant.

Techniques Used: Quantitative RT-PCR, Binding Assay, Luciferase, Reporter Gene Assay, Fluorescence, Transfection

Verification of existence of circ-HIPK3-miR-17-3p-ADCY6 axis. (A) Left: Bar graph showing that miR-17-3p could significantly decrease the fluorescence density of NMCMs with pmirGLO-wt-ADCY6-3'UTR; Right: The binding sites of WT and MUT sequence in ADCY6 3'UTR with miR-17-3p. * p < 0.05. (B) WB showing that miR-17-3p can decrease the level of ACDY6 and inhibitor can increase it. (C) ADCY6 can be up- or down-regulated by pCMV-ADCY6 or si-ADCY6 at genetic and protein level. * p < 0.05. (D) The overexpression of ADCY6 can increase the level of p-RyR2 and p-PLN and vice versa. (E) The ADCY6 overexpression can increase the peak FI of fluo-3 in NMCMs activated by carbamylcholine and vice versa. * p < 0.05, n = 15. (F) Bar graph showing the variation of ADCY6 in NMCMs had little effects on FDHM of Ca 2+ transient. NS = not significant, n = 15. (G) Left: The peak FI of 475nm can be downregulated by ADCY6 overexpression and upregulated by its under-expression; Right: The peak FI of 530nm can be upregulated by ADCY6 overexpression and downregulated by its under-expression. * p < 0.05, n = 15. (H) WB showing the increase of ADCY6 caused by circ-HIPK3 can be attenuated by miR-17-3p and the reduction induced by si-circ-HIPK3 can be rescued by inhibitor.
Figure Legend Snippet: Verification of existence of circ-HIPK3-miR-17-3p-ADCY6 axis. (A) Left: Bar graph showing that miR-17-3p could significantly decrease the fluorescence density of NMCMs with pmirGLO-wt-ADCY6-3'UTR; Right: The binding sites of WT and MUT sequence in ADCY6 3'UTR with miR-17-3p. * p < 0.05. (B) WB showing that miR-17-3p can decrease the level of ACDY6 and inhibitor can increase it. (C) ADCY6 can be up- or down-regulated by pCMV-ADCY6 or si-ADCY6 at genetic and protein level. * p < 0.05. (D) The overexpression of ADCY6 can increase the level of p-RyR2 and p-PLN and vice versa. (E) The ADCY6 overexpression can increase the peak FI of fluo-3 in NMCMs activated by carbamylcholine and vice versa. * p < 0.05, n = 15. (F) Bar graph showing the variation of ADCY6 in NMCMs had little effects on FDHM of Ca 2+ transient. NS = not significant, n = 15. (G) Left: The peak FI of 475nm can be downregulated by ADCY6 overexpression and upregulated by its under-expression; Right: The peak FI of 530nm can be upregulated by ADCY6 overexpression and downregulated by its under-expression. * p < 0.05, n = 15. (H) WB showing the increase of ADCY6 caused by circ-HIPK3 can be attenuated by miR-17-3p and the reduction induced by si-circ-HIPK3 can be rescued by inhibitor.

Techniques Used: Fluorescence, Binding Assay, Sequencing, Over Expression, Expressing

AAV9-shRNA in vivo improved the cardiac function post MI. (A) qRT-PCR showed HIPK3 and circ-HIPK3 increased in NC group and decreased in experiment group respectively. * p<0.05. (B) Left: Representative images of hearts sections by TTC staining (Viable myocardium stained red, and the infarcted areas appeared pale). Dotted box showing the infarcted areas. Right: Bar graph showing percent of infarcted LV in experiment group decreased significantly compared to that in NC group. * p<0.001. (C)(D) Results of echocardiography showed that the cardiac function of heart in experiment group increased significantly compared with that in NC group. * p<0.05. (E) Upper: Masson staining showed the degree of fibrosis of heart in NC group was much higher than that in experiment group, normal group or control group. Lower: WB showed col1 (collagen 1) and col3 (collagen 3) increased significantly in NC group. (F) Immuno-blotting showing that ADCY6 increased in NC group but the level of RyR2, PLN and their phosphorylated form decreased in NC group. (G) QRT-PCR showed that PLN, RyR2 and SERCA2a decreased significantly in NC group when compared to these in experiment group but ADCY6 changed little. * p<0.05, NS = not significant.
Figure Legend Snippet: AAV9-shRNA in vivo improved the cardiac function post MI. (A) qRT-PCR showed HIPK3 and circ-HIPK3 increased in NC group and decreased in experiment group respectively. * p<0.05. (B) Left: Representative images of hearts sections by TTC staining (Viable myocardium stained red, and the infarcted areas appeared pale). Dotted box showing the infarcted areas. Right: Bar graph showing percent of infarcted LV in experiment group decreased significantly compared to that in NC group. * p<0.001. (C)(D) Results of echocardiography showed that the cardiac function of heart in experiment group increased significantly compared with that in NC group. * p<0.05. (E) Upper: Masson staining showed the degree of fibrosis of heart in NC group was much higher than that in experiment group, normal group or control group. Lower: WB showed col1 (collagen 1) and col3 (collagen 3) increased significantly in NC group. (F) Immuno-blotting showing that ADCY6 increased in NC group but the level of RyR2, PLN and their phosphorylated form decreased in NC group. (G) QRT-PCR showed that PLN, RyR2 and SERCA2a decreased significantly in NC group when compared to these in experiment group but ADCY6 changed little. * p<0.05, NS = not significant.

Techniques Used: shRNA, In Vivo, Quantitative RT-PCR, Staining


Structured Review

Abcam anti p ryr2 ser2808
Effect of CHSSC on the expression of CaMKII, p-CaMKII, <t>RyR2,</t> and <t>p-RyR2</t> in rats with myocardial ischemia. (a) Western blot analysis of the expression levels of p-CaMKII, CaMKII, and RyR2. Glyceraldehyde 3-phosphate dehydrogenase was used as the loading control. (b) Quantitative analysis of the relative expression levels of p-CaMKII. (c) Quantitative analysis of the relative expression levels of CaMKII. (d) Quantitative analysis of the relative expression levels of p-RyR2. (e) Quantitative analysis of the relative expression levels of RyR2. N = 3 mice per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the sham operation group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model group.
Anti P Ryr2 Ser2808, supplied by Abcam, 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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Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti p ryr2 ser2808 - by Bioz Stars, 2023-01
86/100 stars

Images

1) Product Images from "Chai-Hu-San-Shen Capsule Ameliorates Ventricular Arrhythmia Through Inhibition of the CaMKII/FKBP12.6/RyR2/Ca 2+ Signaling Pathway in Rats with Myocardial Ischemia"

Article Title: Chai-Hu-San-Shen Capsule Ameliorates Ventricular Arrhythmia Through Inhibition of the CaMKII/FKBP12.6/RyR2/Ca 2+ Signaling Pathway in Rats with Myocardial Ischemia

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

doi: 10.1155/2022/2670473

Effect of CHSSC on the expression of CaMKII, p-CaMKII, RyR2, and p-RyR2 in rats with myocardial ischemia. (a) Western blot analysis of the expression levels of p-CaMKII, CaMKII, and RyR2. Glyceraldehyde 3-phosphate dehydrogenase was used as the loading control. (b) Quantitative analysis of the relative expression levels of p-CaMKII. (c) Quantitative analysis of the relative expression levels of CaMKII. (d) Quantitative analysis of the relative expression levels of p-RyR2. (e) Quantitative analysis of the relative expression levels of RyR2. N = 3 mice per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the sham operation group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model group.
Figure Legend Snippet: Effect of CHSSC on the expression of CaMKII, p-CaMKII, RyR2, and p-RyR2 in rats with myocardial ischemia. (a) Western blot analysis of the expression levels of p-CaMKII, CaMKII, and RyR2. Glyceraldehyde 3-phosphate dehydrogenase was used as the loading control. (b) Quantitative analysis of the relative expression levels of p-CaMKII. (c) Quantitative analysis of the relative expression levels of CaMKII. (d) Quantitative analysis of the relative expression levels of p-RyR2. (e) Quantitative analysis of the relative expression levels of RyR2. N = 3 mice per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the sham operation group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model group.

Techniques Used: Expressing, Western Blot

Effects of CHSSC on the expression of RyR2, p-RyR2 (Ser2808), and p-RyR2 (Ser2814) in ischemic H9C2 cells. (a) Western blot analysis of the expression levels of RyR2, p-RyR2 (Ser2808), and p-RyR2 (Ser2814). Glyceraldehyde 3-phosphate dehydrogenase was used as the loading control. (b) Quantitative analysis of the relative expression levels of RyR2, p-RyR2 (Ser2808), and p-RyR2 (Ser2814) in each group. N = 3 samples per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the control + H-89 group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model + H-89 group.
Figure Legend Snippet: Effects of CHSSC on the expression of RyR2, p-RyR2 (Ser2808), and p-RyR2 (Ser2814) in ischemic H9C2 cells. (a) Western blot analysis of the expression levels of RyR2, p-RyR2 (Ser2808), and p-RyR2 (Ser2814). Glyceraldehyde 3-phosphate dehydrogenase was used as the loading control. (b) Quantitative analysis of the relative expression levels of RyR2, p-RyR2 (Ser2808), and p-RyR2 (Ser2814) in each group. N = 3 samples per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the control + H-89 group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model + H-89 group.

Techniques Used: Expressing, Western Blot

Effect of CHSSC on the interaction between FKBP12.6 and RyR2 in ischemic H9C2 cells. (a) Co-immunoprecipitation was used to evaluate the interaction between FKBP12.6 and RyR2. (b) Quantitative analysis of the level of RyR2/FKBP12.6 in each group. N = 3 samples per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the control + H-89 group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model + H-89 group.
Figure Legend Snippet: Effect of CHSSC on the interaction between FKBP12.6 and RyR2 in ischemic H9C2 cells. (a) Co-immunoprecipitation was used to evaluate the interaction between FKBP12.6 and RyR2. (b) Quantitative analysis of the level of RyR2/FKBP12.6 in each group. N = 3 samples per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the control + H-89 group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model + H-89 group.

Techniques Used: Immunoprecipitation


Structured Review

Abcam anti p ryanodine receptor 2 ps2808
PKA pathway phosphoproteomic changes in STK25 −/− compared with wild type
Anti P Ryanodine Receptor 2 Ps2808, supplied by Abcam, 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 p ryanodine receptor 2 ps2808/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti p ryanodine receptor 2 ps2808 - by Bioz Stars, 2023-01
86/100 stars

Images

1) Product Images from "STK25 inhibits PKA signaling by phosphorylating PRKAR1A"

Article Title: STK25 inhibits PKA signaling by phosphorylating PRKAR1A

Journal: Cell reports

doi: 10.1016/j.celrep.2022.111203

PKA pathway phosphoproteomic changes in STK25 −/− compared with wild type
Figure Legend Snippet: PKA pathway phosphoproteomic changes in STK25 −/− compared with wild type

Techniques Used: Expressing

KEY RESOURCES TABLE
Figure Legend Snippet: KEY RESOURCES TABLE

Techniques Used: Recombinant, SYBR Green Assay, In Vitro, Activity Assay, Viability Assay, Sequencing, esiRNA, CRISPR, Knock-Out, Plasmid Preparation, Software


Structured Review

Abcam anti p ryr2
A Target binding of miR-143-3p and NPY. B Dual-luciferase reporter assay for miR-143-3p and NPY. C , D In vivo and in vitro experimental verification of mmu_circ_0000021, expression of miR-143-3p and NPY. E Gene and protein expression of NPY after plasmid transfection. F Western blot showing that the increase of NPY caused by mmu_circ_0000021 was attenuated by miR-143-3p. G Overexpression of NPY increased the level of <t>p-RyR2</t> and p-PLN and vice versa. H Overexpression of NPY increased the fluo-4am value in NMCMs. * P < 0.05; n = 8/group. I Bar graph showing that changes in NPY in NMCMs had little effect on Ca 2+ transients. * P < 0.05, *** P < 0.001 vs. indicated group; NS not significant, n = 8/group.
Anti P Ryr2, supplied by Abcam, 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 p ryr2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti p ryr2 - by Bioz Stars, 2023-01
86/100 stars

Images

1) Product Images from "CircRNA mmu_circ_0000021 regulates microvascular function via the miR-143-3p/NPY axis and intracellular calcium following ischemia/reperfusion injury"

Article Title: CircRNA mmu_circ_0000021 regulates microvascular function via the miR-143-3p/NPY axis and intracellular calcium following ischemia/reperfusion injury

Journal: Cell Death Discovery

doi: 10.1038/s41420-022-01108-z

A Target binding of miR-143-3p and NPY. B Dual-luciferase reporter assay for miR-143-3p and NPY. C , D In vivo and in vitro experimental verification of mmu_circ_0000021, expression of miR-143-3p and NPY. E Gene and protein expression of NPY after plasmid transfection. F Western blot showing that the increase of NPY caused by mmu_circ_0000021 was attenuated by miR-143-3p. G Overexpression of NPY increased the level of p-RyR2 and p-PLN and vice versa. H Overexpression of NPY increased the fluo-4am value in NMCMs. * P < 0.05; n = 8/group. I Bar graph showing that changes in NPY in NMCMs had little effect on Ca 2+ transients. * P < 0.05, *** P < 0.001 vs. indicated group; NS not significant, n = 8/group.
Figure Legend Snippet: A Target binding of miR-143-3p and NPY. B Dual-luciferase reporter assay for miR-143-3p and NPY. C , D In vivo and in vitro experimental verification of mmu_circ_0000021, expression of miR-143-3p and NPY. E Gene and protein expression of NPY after plasmid transfection. F Western blot showing that the increase of NPY caused by mmu_circ_0000021 was attenuated by miR-143-3p. G Overexpression of NPY increased the level of p-RyR2 and p-PLN and vice versa. H Overexpression of NPY increased the fluo-4am value in NMCMs. * P < 0.05; n = 8/group. I Bar graph showing that changes in NPY in NMCMs had little effect on Ca 2+ transients. * P < 0.05, *** P < 0.001 vs. indicated group; NS not significant, n = 8/group.

Techniques Used: Binding Assay, Luciferase, Reporter Assay, In Vivo, In Vitro, Expressing, Plasmid Preparation, Transfection, Western Blot, Over Expression

A , D , E LV function was determined by echocardiograms ( A ) including EF ( D ) and FS ( E ). B , F The size of the infarct and the region of the myocardial infarction were determined using TTC. C In the sham group, the amount of RyR2, PLN, and their phosphorylated forms were reduced. G – L qRT-PCR showed changes in PLN, RyR2, SERCA2a and the changes of miR-143-3p/NPY axis after the inhibition of mmu_circ_0000021 expression. * P < 0.05, ** P < 0.01, vs. indicated group; n = 8/group.
Figure Legend Snippet: A , D , E LV function was determined by echocardiograms ( A ) including EF ( D ) and FS ( E ). B , F The size of the infarct and the region of the myocardial infarction were determined using TTC. C In the sham group, the amount of RyR2, PLN, and their phosphorylated forms were reduced. G – L qRT-PCR showed changes in PLN, RyR2, SERCA2a and the changes of miR-143-3p/NPY axis after the inhibition of mmu_circ_0000021 expression. * P < 0.05, ** P < 0.01, vs. indicated group; n = 8/group.

Techniques Used: Quantitative RT-PCR, Inhibition, Expressing


Structured Review

Abcam anti p ryr2
( A ) Representative immunoblots showing total IP3R1 expression, phosphorylation of serine/tyrosine kinases, and AKAP9 binding to the channels, as assessed by immunoprecipitation (IP: IP3R1); <t>RyR2</t> phosphorylation levels of PKA-induced RyR2 phosphorylation on serine 2808 (pSer-2808); and p-MLC20 levels in aortic tissues from patients with HF ( n = 5) and controls ( n = 4). ( B – G ) Quantification of the immunoblots shown in A . Individual values with the mean ± SEM are shown. * P < 0.05 versus control, by 2-tailed Student’s t test.
Anti P Ryr2, supplied by Abcam, 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 p ryr2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti p ryr2 - by Bioz Stars, 2023-01
86/100 stars

Images

1) Product Images from "IP3 receptor orchestrates maladaptive vascular responses in heart failure"

Article Title: IP3 receptor orchestrates maladaptive vascular responses in heart failure

Journal: The Journal of Clinical Investigation

doi: 10.1172/JCI152859

( A ) Representative immunoblots showing total IP3R1 expression, phosphorylation of serine/tyrosine kinases, and AKAP9 binding to the channels, as assessed by immunoprecipitation (IP: IP3R1); RyR2 phosphorylation levels of PKA-induced RyR2 phosphorylation on serine 2808 (pSer-2808); and p-MLC20 levels in aortic tissues from patients with HF ( n = 5) and controls ( n = 4). ( B – G ) Quantification of the immunoblots shown in A . Individual values with the mean ± SEM are shown. * P < 0.05 versus control, by 2-tailed Student’s t test.
Figure Legend Snippet: ( A ) Representative immunoblots showing total IP3R1 expression, phosphorylation of serine/tyrosine kinases, and AKAP9 binding to the channels, as assessed by immunoprecipitation (IP: IP3R1); RyR2 phosphorylation levels of PKA-induced RyR2 phosphorylation on serine 2808 (pSer-2808); and p-MLC20 levels in aortic tissues from patients with HF ( n = 5) and controls ( n = 4). ( B – G ) Quantification of the immunoblots shown in A . Individual values with the mean ± SEM are shown. * P < 0.05 versus control, by 2-tailed Student’s t test.

Techniques Used: Western Blot, Expressing, Binding Assay, Immunoprecipitation

ATII binds to GPCRs (G q –G 11 ) and activates PLC, which hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP2), resulting in 2 second messengers: IP3 and diacylglycerol (DAG). IP3 binds to its receptor IP3R1 on the SR, causing Ca 2+ release into the cytosol. Furthermore, increased catecholamines during HF bind to adrenergic (AD) receptors and activate the Gs protein and AC, leading to increased levels of cAMP. cAMP activates PKA, which phosphorylate the IP3R1 channels, causing further Ca 2+ release into the cytosol. IP3R1 binds AKAP9, which anchors a pool of PKA to the channel. Of note, PKA phosphorylates RyR2 and SR/ER Ca 2+ -ATPase type 2 (SERCA2a), which play a role in SR Ca 2+ release and uptake processes, respectively. An increase in the cytosolic Ca 2+ concentration activates the CAM protein. CAM activates MLCK, which in turn phosphorylates MLC20, leading to smooth muscle contraction and vasoconstriction. Chronic vasoconstriction increases cardiac afterload, thereby promoting decompensated HF. Both genetic depletion of VSMC IP3R1 and pharmacologic inhibition of MLCK with ML-7 attenuate MLCK activation and phosphorylation of MLC20, thus reducing vasoconstriction and cardiac afterload in failing hearts. MLCP, myosin light chain phosphatase; PLN, phospholamban.
Figure Legend Snippet: ATII binds to GPCRs (G q –G 11 ) and activates PLC, which hydrolyzes phosphatidylinositol 4,5-bisphosphate (PIP2), resulting in 2 second messengers: IP3 and diacylglycerol (DAG). IP3 binds to its receptor IP3R1 on the SR, causing Ca 2+ release into the cytosol. Furthermore, increased catecholamines during HF bind to adrenergic (AD) receptors and activate the Gs protein and AC, leading to increased levels of cAMP. cAMP activates PKA, which phosphorylate the IP3R1 channels, causing further Ca 2+ release into the cytosol. IP3R1 binds AKAP9, which anchors a pool of PKA to the channel. Of note, PKA phosphorylates RyR2 and SR/ER Ca 2+ -ATPase type 2 (SERCA2a), which play a role in SR Ca 2+ release and uptake processes, respectively. An increase in the cytosolic Ca 2+ concentration activates the CAM protein. CAM activates MLCK, which in turn phosphorylates MLC20, leading to smooth muscle contraction and vasoconstriction. Chronic vasoconstriction increases cardiac afterload, thereby promoting decompensated HF. Both genetic depletion of VSMC IP3R1 and pharmacologic inhibition of MLCK with ML-7 attenuate MLCK activation and phosphorylation of MLC20, thus reducing vasoconstriction and cardiac afterload in failing hearts. MLCP, myosin light chain phosphatase; PLN, phospholamban.

Techniques Used: Concentration Assay, Inhibition, Activation Assay


Structured Review

Abcam anti p ryanodine receptor 2 s2808
A) Immunoblot of PRKAR1A phosphorylation sites S77 and S83 in STK25 +/+ and STK25 -/- cardiomyocyte protein lysates. B) Immunoblots of phosphorylation sites S77 and S83 of PRKAR1A in STK25 -/- cardiomyocytes transfected with empty vector (EV), wild type STK25 and kinase dead K49R/T174A (loaded in duplicate). C) Forskolin (10μM) stimulated STK25 +/+ and STK25 -/- cardiomyocytes immunoblotted for phosphorylation of PRKAR1A and <t>RYR2.</t> D) Immunoprecipitation of Flag-STK25 expressed in HEK293T cells and immunoblotted for PRKAR1A and GM130 (positive control binding partner). E) Co-immunoprecipitation of PRKAR1A-V5 with STK25 and PRKACA in HEK293T cells treated with forskolin (10μM). F) In vitro kinase assay of purified STK25 and PRKAR1A, immunoblotted for phosphorylation of S83 of PRKAR1A.
Anti P Ryanodine Receptor 2 S2808, supplied by Abcam, 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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Price from $9.99 to $1999.99
anti p ryanodine receptor 2 s2808 - by Bioz Stars, 2023-01
86/100 stars

Images

1) Product Images from "STK25 inhibits PKA signaling by phosphorylating PRKAR1A"

Article Title: STK25 inhibits PKA signaling by phosphorylating PRKAR1A

Journal: bioRxiv

doi: 10.1101/2022.01.27.478084

A) Immunoblot of PRKAR1A phosphorylation sites S77 and S83 in STK25 +/+ and STK25 -/- cardiomyocyte protein lysates. B) Immunoblots of phosphorylation sites S77 and S83 of PRKAR1A in STK25 -/- cardiomyocytes transfected with empty vector (EV), wild type STK25 and kinase dead K49R/T174A (loaded in duplicate). C) Forskolin (10μM) stimulated STK25 +/+ and STK25 -/- cardiomyocytes immunoblotted for phosphorylation of PRKAR1A and RYR2. D) Immunoprecipitation of Flag-STK25 expressed in HEK293T cells and immunoblotted for PRKAR1A and GM130 (positive control binding partner). E) Co-immunoprecipitation of PRKAR1A-V5 with STK25 and PRKACA in HEK293T cells treated with forskolin (10μM). F) In vitro kinase assay of purified STK25 and PRKAR1A, immunoblotted for phosphorylation of S83 of PRKAR1A.
Figure Legend Snippet: A) Immunoblot of PRKAR1A phosphorylation sites S77 and S83 in STK25 +/+ and STK25 -/- cardiomyocyte protein lysates. B) Immunoblots of phosphorylation sites S77 and S83 of PRKAR1A in STK25 -/- cardiomyocytes transfected with empty vector (EV), wild type STK25 and kinase dead K49R/T174A (loaded in duplicate). C) Forskolin (10μM) stimulated STK25 +/+ and STK25 -/- cardiomyocytes immunoblotted for phosphorylation of PRKAR1A and RYR2. D) Immunoprecipitation of Flag-STK25 expressed in HEK293T cells and immunoblotted for PRKAR1A and GM130 (positive control binding partner). E) Co-immunoprecipitation of PRKAR1A-V5 with STK25 and PRKACA in HEK293T cells treated with forskolin (10μM). F) In vitro kinase assay of purified STK25 and PRKAR1A, immunoblotted for phosphorylation of S83 of PRKAR1A.

Techniques Used: Western Blot, Transfection, Plasmid Preparation, Immunoprecipitation, Positive Control, Binding Assay, In Vitro, Kinase Assay, Purification


Structured Review

Abcam anti p ryr2
Anti P Ryr2, supplied by Abcam, 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 p ryr2/product/Abcam
Average 86 stars, based on 1 article reviews
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Structured Review

Abcam anti p ryr2
Anti P Ryr2, supplied by Abcam, 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 p ryr2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti p ryr2 - by Bioz Stars, 2023-01
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Structured Review

Abcam p ryr2
Fgf21 protected against structural and electrical remodeling in the atria. (A) Immunofluorescence analysis was performed to analyze myosin heavy chain (MHC). (B) Summary of the relative fluorescence intensity of MHC in each group of HL-1 atrial cells. (C) The expression of Troponin I, CACNA1C, Calpain, <t>p-RyR2,</t> and <t>t-RyR2</t> at protein levels in each group of HL-1 atrial cells. * p < 0.05.
P Ryr2, supplied by Abcam, 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/p ryr2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
p ryr2 - by Bioz Stars, 2023-01
86/100 stars

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1) Product Images from "Fibroblast Growth Factor 21 Protects Against Atrial Remodeling via Reducing Oxidative Stress"

Article Title: Fibroblast Growth Factor 21 Protects Against Atrial Remodeling via Reducing Oxidative Stress

Journal: Frontiers in Cardiovascular Medicine

doi: 10.3389/fcvm.2021.720581

Fgf21 protected against structural and electrical remodeling in the atria. (A) Immunofluorescence analysis was performed to analyze myosin heavy chain (MHC). (B) Summary of the relative fluorescence intensity of MHC in each group of HL-1 atrial cells. (C) The expression of Troponin I, CACNA1C, Calpain, p-RyR2, and t-RyR2 at protein levels in each group of HL-1 atrial cells. * p < 0.05.
Figure Legend Snippet: Fgf21 protected against structural and electrical remodeling in the atria. (A) Immunofluorescence analysis was performed to analyze myosin heavy chain (MHC). (B) Summary of the relative fluorescence intensity of MHC in each group of HL-1 atrial cells. (C) The expression of Troponin I, CACNA1C, Calpain, p-RyR2, and t-RyR2 at protein levels in each group of HL-1 atrial cells. * p < 0.05.

Techniques Used: Immunofluorescence, Fluorescence, Expressing

Nuclear factor erythroid 2-related factor 2 (Nrf2) positively regulated anti-oxidative genes expression induced by Fgf21. (A) Western blot was conducted to evaluate the expression of SOD2 and UCP3 in HL-1 atrial cells by knockdown of Nrf2 with si-Nrf2. (B) Western blot was conducted to evaluate the expression of Troponin I, Tgf-β, ox-CaMKII, t-CaMKII, CACNA1C, Calpain, p-RyR2, and t-RyR2 in HL-1 atrial cells by knockdown of Nrf2 with si-Nrf2. * p < 0.05.
Figure Legend Snippet: Nuclear factor erythroid 2-related factor 2 (Nrf2) positively regulated anti-oxidative genes expression induced by Fgf21. (A) Western blot was conducted to evaluate the expression of SOD2 and UCP3 in HL-1 atrial cells by knockdown of Nrf2 with si-Nrf2. (B) Western blot was conducted to evaluate the expression of Troponin I, Tgf-β, ox-CaMKII, t-CaMKII, CACNA1C, Calpain, p-RyR2, and t-RyR2 in HL-1 atrial cells by knockdown of Nrf2 with si-Nrf2. * p < 0.05.

Techniques Used: Expressing, Western Blot


Structured Review

Abcam anti phosphorylated p ryr 2
Anti Phosphorylated P Ryr 2, supplied by Abcam, 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 phosphorylated p ryr 2/product/Abcam
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
anti phosphorylated p ryr 2 - by Bioz Stars, 2023-01
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  • 86
    Abcam anti p ryr2
    MiR-17-3p was the downstream target of circ-HIPK3. (A) Results of WB showing that <t>p-RyR2</t> and p-PLN increased in circ-HIPK3 overexpressed NMCMs and vice versa. (B) QRT-PCR showing that PLN and <t>RyR2</t> changed little in circ-HIPK3 over- or under- expressed NMCMs. NS = not significant. (C) Schematic image showing that seven possible binding sites of miR-17-3p to circ-HIPK3 were found. (D) Dual-luciferase reporter gene assay showed miR-17-3p could decrease the fluorescence density of NMCMs transfected with pmirGLO- wt -circ-HIPK3. * p<0.05 . (E) FISH assay showed circ-HIPK3 (red) can interact with miR-17-3p (green), which mainly were around the nucleus (blue). (F) The level of p-RyR2 and p-PLN were downregulated by miR-17-3p mimic or upregulated by inhibitor. (G) The peak FI of fluo-3 decreased in miR-17-3p transfected NMCMs and increased in inhibitor transfected NMCMs. ** p < 0.01, n = 15. (H) The FDHM of NMCMs activated by carbamylcholine was not affected by miR-17-3p or inhibitor. NS = not significant, n = 15. (I) The miR-17-3p can decrease the FI of 530nm and increase the 475nm in NMCMs; Inhibitor can increase the FI of 530 and decrease the 475nm. * p < 0.05, n = 15. (J) Both miR-17-3p and inhibitor had no effects on PLN and RyR2 in genetic level. NS = not significant.
    Anti P Ryr2, supplied by Abcam, 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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    Abcam anti p ryr2 ser2808
    Effect of CHSSC on the expression of CaMKII, p-CaMKII, <t>RyR2,</t> and <t>p-RyR2</t> in rats with myocardial ischemia. (a) Western blot analysis of the expression levels of p-CaMKII, CaMKII, and RyR2. Glyceraldehyde 3-phosphate dehydrogenase was used as the loading control. (b) Quantitative analysis of the relative expression levels of p-CaMKII. (c) Quantitative analysis of the relative expression levels of CaMKII. (d) Quantitative analysis of the relative expression levels of p-RyR2. (e) Quantitative analysis of the relative expression levels of RyR2. N = 3 mice per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the sham operation group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model group.
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    Abcam anti p ryanodine receptor 2 ps2808
    PKA pathway phosphoproteomic changes in STK25 −/− compared with wild type
    Anti P Ryanodine Receptor 2 Ps2808, supplied by Abcam, 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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    Abcam anti p ryanodine receptor 2 s2808
    A) Immunoblot of PRKAR1A phosphorylation sites S77 and S83 in STK25 +/+ and STK25 -/- cardiomyocyte protein lysates. B) Immunoblots of phosphorylation sites S77 and S83 of PRKAR1A in STK25 -/- cardiomyocytes transfected with empty vector (EV), wild type STK25 and kinase dead K49R/T174A (loaded in duplicate). C) Forskolin (10μM) stimulated STK25 +/+ and STK25 -/- cardiomyocytes immunoblotted for phosphorylation of PRKAR1A and <t>RYR2.</t> D) Immunoprecipitation of Flag-STK25 expressed in HEK293T cells and immunoblotted for PRKAR1A and GM130 (positive control binding partner). E) Co-immunoprecipitation of PRKAR1A-V5 with STK25 and PRKACA in HEK293T cells treated with forskolin (10μM). F) In vitro kinase assay of purified STK25 and PRKAR1A, immunoblotted for phosphorylation of S83 of PRKAR1A.
    Anti P Ryanodine Receptor 2 S2808, supplied by Abcam, 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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    Abcam p ryr2
    Fgf21 protected against structural and electrical remodeling in the atria. (A) Immunofluorescence analysis was performed to analyze myosin heavy chain (MHC). (B) Summary of the relative fluorescence intensity of MHC in each group of HL-1 atrial cells. (C) The expression of Troponin I, CACNA1C, Calpain, <t>p-RyR2,</t> and <t>t-RyR2</t> at protein levels in each group of HL-1 atrial cells. * p < 0.05.
    P Ryr2, supplied by Abcam, 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/p ryr2/product/Abcam
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    Abcam anti phosphorylated p ryr 2
    Fgf21 protected against structural and electrical remodeling in the atria. (A) Immunofluorescence analysis was performed to analyze myosin heavy chain (MHC). (B) Summary of the relative fluorescence intensity of MHC in each group of HL-1 atrial cells. (C) The expression of Troponin I, CACNA1C, Calpain, <t>p-RyR2,</t> and <t>t-RyR2</t> at protein levels in each group of HL-1 atrial cells. * p < 0.05.
    Anti Phosphorylated P Ryr 2, supplied by Abcam, 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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    MiR-17-3p was the downstream target of circ-HIPK3. (A) Results of WB showing that p-RyR2 and p-PLN increased in circ-HIPK3 overexpressed NMCMs and vice versa. (B) QRT-PCR showing that PLN and RyR2 changed little in circ-HIPK3 over- or under- expressed NMCMs. NS = not significant. (C) Schematic image showing that seven possible binding sites of miR-17-3p to circ-HIPK3 were found. (D) Dual-luciferase reporter gene assay showed miR-17-3p could decrease the fluorescence density of NMCMs transfected with pmirGLO- wt -circ-HIPK3. * p<0.05 . (E) FISH assay showed circ-HIPK3 (red) can interact with miR-17-3p (green), which mainly were around the nucleus (blue). (F) The level of p-RyR2 and p-PLN were downregulated by miR-17-3p mimic or upregulated by inhibitor. (G) The peak FI of fluo-3 decreased in miR-17-3p transfected NMCMs and increased in inhibitor transfected NMCMs. ** p < 0.01, n = 15. (H) The FDHM of NMCMs activated by carbamylcholine was not affected by miR-17-3p or inhibitor. NS = not significant, n = 15. (I) The miR-17-3p can decrease the FI of 530nm and increase the 475nm in NMCMs; Inhibitor can increase the FI of 530 and decrease the 475nm. * p < 0.05, n = 15. (J) Both miR-17-3p and inhibitor had no effects on PLN and RyR2 in genetic level. NS = not significant.

    Journal: International Journal of Biological Sciences

    Article Title: Circ-HIPK3 Strengthens the Effects of Adrenaline in Heart Failure by MiR-17-3p - ADCY6 Axis

    doi: 10.7150/ijbs.36149

    Figure Lengend Snippet: MiR-17-3p was the downstream target of circ-HIPK3. (A) Results of WB showing that p-RyR2 and p-PLN increased in circ-HIPK3 overexpressed NMCMs and vice versa. (B) QRT-PCR showing that PLN and RyR2 changed little in circ-HIPK3 over- or under- expressed NMCMs. NS = not significant. (C) Schematic image showing that seven possible binding sites of miR-17-3p to circ-HIPK3 were found. (D) Dual-luciferase reporter gene assay showed miR-17-3p could decrease the fluorescence density of NMCMs transfected with pmirGLO- wt -circ-HIPK3. * p<0.05 . (E) FISH assay showed circ-HIPK3 (red) can interact with miR-17-3p (green), which mainly were around the nucleus (blue). (F) The level of p-RyR2 and p-PLN were downregulated by miR-17-3p mimic or upregulated by inhibitor. (G) The peak FI of fluo-3 decreased in miR-17-3p transfected NMCMs and increased in inhibitor transfected NMCMs. ** p < 0.01, n = 15. (H) The FDHM of NMCMs activated by carbamylcholine was not affected by miR-17-3p or inhibitor. NS = not significant, n = 15. (I) The miR-17-3p can decrease the FI of 530nm and increase the 475nm in NMCMs; Inhibitor can increase the FI of 530 and decrease the 475nm. * p < 0.05, n = 15. (J) Both miR-17-3p and inhibitor had no effects on PLN and RyR2 in genetic level. NS = not significant.

    Article Snippet: Anti-RyR2, anti-PLN, anti-p-PLN, anti-p-RyR2, anti-His were purchased from Abcam (Britain).

    Techniques: Quantitative RT-PCR, Binding Assay, Luciferase, Reporter Gene Assay, Fluorescence, Transfection

    Verification of existence of circ-HIPK3-miR-17-3p-ADCY6 axis. (A) Left: Bar graph showing that miR-17-3p could significantly decrease the fluorescence density of NMCMs with pmirGLO-wt-ADCY6-3'UTR; Right: The binding sites of WT and MUT sequence in ADCY6 3'UTR with miR-17-3p. * p < 0.05. (B) WB showing that miR-17-3p can decrease the level of ACDY6 and inhibitor can increase it. (C) ADCY6 can be up- or down-regulated by pCMV-ADCY6 or si-ADCY6 at genetic and protein level. * p < 0.05. (D) The overexpression of ADCY6 can increase the level of p-RyR2 and p-PLN and vice versa. (E) The ADCY6 overexpression can increase the peak FI of fluo-3 in NMCMs activated by carbamylcholine and vice versa. * p < 0.05, n = 15. (F) Bar graph showing the variation of ADCY6 in NMCMs had little effects on FDHM of Ca 2+ transient. NS = not significant, n = 15. (G) Left: The peak FI of 475nm can be downregulated by ADCY6 overexpression and upregulated by its under-expression; Right: The peak FI of 530nm can be upregulated by ADCY6 overexpression and downregulated by its under-expression. * p < 0.05, n = 15. (H) WB showing the increase of ADCY6 caused by circ-HIPK3 can be attenuated by miR-17-3p and the reduction induced by si-circ-HIPK3 can be rescued by inhibitor.

    Journal: International Journal of Biological Sciences

    Article Title: Circ-HIPK3 Strengthens the Effects of Adrenaline in Heart Failure by MiR-17-3p - ADCY6 Axis

    doi: 10.7150/ijbs.36149

    Figure Lengend Snippet: Verification of existence of circ-HIPK3-miR-17-3p-ADCY6 axis. (A) Left: Bar graph showing that miR-17-3p could significantly decrease the fluorescence density of NMCMs with pmirGLO-wt-ADCY6-3'UTR; Right: The binding sites of WT and MUT sequence in ADCY6 3'UTR with miR-17-3p. * p < 0.05. (B) WB showing that miR-17-3p can decrease the level of ACDY6 and inhibitor can increase it. (C) ADCY6 can be up- or down-regulated by pCMV-ADCY6 or si-ADCY6 at genetic and protein level. * p < 0.05. (D) The overexpression of ADCY6 can increase the level of p-RyR2 and p-PLN and vice versa. (E) The ADCY6 overexpression can increase the peak FI of fluo-3 in NMCMs activated by carbamylcholine and vice versa. * p < 0.05, n = 15. (F) Bar graph showing the variation of ADCY6 in NMCMs had little effects on FDHM of Ca 2+ transient. NS = not significant, n = 15. (G) Left: The peak FI of 475nm can be downregulated by ADCY6 overexpression and upregulated by its under-expression; Right: The peak FI of 530nm can be upregulated by ADCY6 overexpression and downregulated by its under-expression. * p < 0.05, n = 15. (H) WB showing the increase of ADCY6 caused by circ-HIPK3 can be attenuated by miR-17-3p and the reduction induced by si-circ-HIPK3 can be rescued by inhibitor.

    Article Snippet: Anti-RyR2, anti-PLN, anti-p-PLN, anti-p-RyR2, anti-His were purchased from Abcam (Britain).

    Techniques: Fluorescence, Binding Assay, Sequencing, Over Expression, Expressing

    AAV9-shRNA in vivo improved the cardiac function post MI. (A) qRT-PCR showed HIPK3 and circ-HIPK3 increased in NC group and decreased in experiment group respectively. * p<0.05. (B) Left: Representative images of hearts sections by TTC staining (Viable myocardium stained red, and the infarcted areas appeared pale). Dotted box showing the infarcted areas. Right: Bar graph showing percent of infarcted LV in experiment group decreased significantly compared to that in NC group. * p<0.001. (C)(D) Results of echocardiography showed that the cardiac function of heart in experiment group increased significantly compared with that in NC group. * p<0.05. (E) Upper: Masson staining showed the degree of fibrosis of heart in NC group was much higher than that in experiment group, normal group or control group. Lower: WB showed col1 (collagen 1) and col3 (collagen 3) increased significantly in NC group. (F) Immuno-blotting showing that ADCY6 increased in NC group but the level of RyR2, PLN and their phosphorylated form decreased in NC group. (G) QRT-PCR showed that PLN, RyR2 and SERCA2a decreased significantly in NC group when compared to these in experiment group but ADCY6 changed little. * p<0.05, NS = not significant.

    Journal: International Journal of Biological Sciences

    Article Title: Circ-HIPK3 Strengthens the Effects of Adrenaline in Heart Failure by MiR-17-3p - ADCY6 Axis

    doi: 10.7150/ijbs.36149

    Figure Lengend Snippet: AAV9-shRNA in vivo improved the cardiac function post MI. (A) qRT-PCR showed HIPK3 and circ-HIPK3 increased in NC group and decreased in experiment group respectively. * p<0.05. (B) Left: Representative images of hearts sections by TTC staining (Viable myocardium stained red, and the infarcted areas appeared pale). Dotted box showing the infarcted areas. Right: Bar graph showing percent of infarcted LV in experiment group decreased significantly compared to that in NC group. * p<0.001. (C)(D) Results of echocardiography showed that the cardiac function of heart in experiment group increased significantly compared with that in NC group. * p<0.05. (E) Upper: Masson staining showed the degree of fibrosis of heart in NC group was much higher than that in experiment group, normal group or control group. Lower: WB showed col1 (collagen 1) and col3 (collagen 3) increased significantly in NC group. (F) Immuno-blotting showing that ADCY6 increased in NC group but the level of RyR2, PLN and their phosphorylated form decreased in NC group. (G) QRT-PCR showed that PLN, RyR2 and SERCA2a decreased significantly in NC group when compared to these in experiment group but ADCY6 changed little. * p<0.05, NS = not significant.

    Article Snippet: Anti-RyR2, anti-PLN, anti-p-PLN, anti-p-RyR2, anti-His were purchased from Abcam (Britain).

    Techniques: shRNA, In Vivo, Quantitative RT-PCR, Staining

    Effect of CHSSC on the expression of CaMKII, p-CaMKII, RyR2, and p-RyR2 in rats with myocardial ischemia. (a) Western blot analysis of the expression levels of p-CaMKII, CaMKII, and RyR2. Glyceraldehyde 3-phosphate dehydrogenase was used as the loading control. (b) Quantitative analysis of the relative expression levels of p-CaMKII. (c) Quantitative analysis of the relative expression levels of CaMKII. (d) Quantitative analysis of the relative expression levels of p-RyR2. (e) Quantitative analysis of the relative expression levels of RyR2. N = 3 mice per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the sham operation group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model group.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Chai-Hu-San-Shen Capsule Ameliorates Ventricular Arrhythmia Through Inhibition of the CaMKII/FKBP12.6/RyR2/Ca 2+ Signaling Pathway in Rats with Myocardial Ischemia

    doi: 10.1155/2022/2670473

    Figure Lengend Snippet: Effect of CHSSC on the expression of CaMKII, p-CaMKII, RyR2, and p-RyR2 in rats with myocardial ischemia. (a) Western blot analysis of the expression levels of p-CaMKII, CaMKII, and RyR2. Glyceraldehyde 3-phosphate dehydrogenase was used as the loading control. (b) Quantitative analysis of the relative expression levels of p-CaMKII. (c) Quantitative analysis of the relative expression levels of CaMKII. (d) Quantitative analysis of the relative expression levels of p-RyR2. (e) Quantitative analysis of the relative expression levels of RyR2. N = 3 mice per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the sham operation group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model group.

    Article Snippet: Anti-RyR2 (#ab2868) and anti-p-RyR2 (Ser2808) (#ab59225) were purchased from Abcam (Cambridge, UK).

    Techniques: Expressing, Western Blot

    Effects of CHSSC on the expression of RyR2, p-RyR2 (Ser2808), and p-RyR2 (Ser2814) in ischemic H9C2 cells. (a) Western blot analysis of the expression levels of RyR2, p-RyR2 (Ser2808), and p-RyR2 (Ser2814). Glyceraldehyde 3-phosphate dehydrogenase was used as the loading control. (b) Quantitative analysis of the relative expression levels of RyR2, p-RyR2 (Ser2808), and p-RyR2 (Ser2814) in each group. N = 3 samples per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the control + H-89 group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model + H-89 group.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Chai-Hu-San-Shen Capsule Ameliorates Ventricular Arrhythmia Through Inhibition of the CaMKII/FKBP12.6/RyR2/Ca 2+ Signaling Pathway in Rats with Myocardial Ischemia

    doi: 10.1155/2022/2670473

    Figure Lengend Snippet: Effects of CHSSC on the expression of RyR2, p-RyR2 (Ser2808), and p-RyR2 (Ser2814) in ischemic H9C2 cells. (a) Western blot analysis of the expression levels of RyR2, p-RyR2 (Ser2808), and p-RyR2 (Ser2814). Glyceraldehyde 3-phosphate dehydrogenase was used as the loading control. (b) Quantitative analysis of the relative expression levels of RyR2, p-RyR2 (Ser2808), and p-RyR2 (Ser2814) in each group. N = 3 samples per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the control + H-89 group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model + H-89 group.

    Article Snippet: Anti-RyR2 (#ab2868) and anti-p-RyR2 (Ser2808) (#ab59225) were purchased from Abcam (Cambridge, UK).

    Techniques: Expressing, Western Blot

    Effect of CHSSC on the interaction between FKBP12.6 and RyR2 in ischemic H9C2 cells. (a) Co-immunoprecipitation was used to evaluate the interaction between FKBP12.6 and RyR2. (b) Quantitative analysis of the level of RyR2/FKBP12.6 in each group. N = 3 samples per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the control + H-89 group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model + H-89 group.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Chai-Hu-San-Shen Capsule Ameliorates Ventricular Arrhythmia Through Inhibition of the CaMKII/FKBP12.6/RyR2/Ca 2+ Signaling Pathway in Rats with Myocardial Ischemia

    doi: 10.1155/2022/2670473

    Figure Lengend Snippet: Effect of CHSSC on the interaction between FKBP12.6 and RyR2 in ischemic H9C2 cells. (a) Co-immunoprecipitation was used to evaluate the interaction between FKBP12.6 and RyR2. (b) Quantitative analysis of the level of RyR2/FKBP12.6 in each group. N = 3 samples per group. All data are expressed as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 vs. the control + H-89 group; # P < 0.05, ## P < 0.05, ### P < 0.001 vs. the model + H-89 group.

    Article Snippet: Anti-RyR2 (#ab2868) and anti-p-RyR2 (Ser2808) (#ab59225) were purchased from Abcam (Cambridge, UK).

    Techniques: Immunoprecipitation

    PKA pathway phosphoproteomic changes in STK25 −/− compared with wild type

    Journal: Cell reports

    Article Title: STK25 inhibits PKA signaling by phosphorylating PRKAR1A

    doi: 10.1016/j.celrep.2022.111203

    Figure Lengend Snippet: PKA pathway phosphoproteomic changes in STK25 −/− compared with wild type

    Article Snippet: For Western blotting antibodies include: HRP conjugated anti-GAPDH (Cell Signaling Technology Cat# 3683, RRID:AB_1642205), anti-STK25 (Abcam Cat# ab157188, RRID:AB_2725788), anti-Flag (Sigma-Aldrich Cat# F3165, RRID:AB_259529), anti-GM130 (Cell Signaling Technology Cat# 12480, RRID:AB_2797933), anti-PRKAR1A (Abcam Cat# ab139695, RRID:AB_2893184), anti-pS77 PRKAR1A (Abcam Cat#ab139682, RRID:AB_2904566), anti-pS83 PRKAR1A (Abcam Cat#ab154851, RRID:AB_2904567), anti-PRKAR2A (ProteinTech Cat# 10142-2-AP), anti-phospholamban (Cell Signaling Technology Cat# 14562, RRID:AB_2798511), anti-phospho-phospholamban -pS16/T17 (Cell Signaling Technology Cat# 8496, RRID:AB_10949102), anti-Ryanodine receptor 2 (Abcam Cat#ab196355, RRID:AB_2904568), anti-p-Ryanodine receptor 2-pS2808 (Abcam Cat# ab59225, RRID:AB_946327), anti-TnI (Cell Signaling Cat# 4002), anti-TnI-pS23/S24 (Cell Signaling Cat# 4004), anti-V5 (Sigma-Aldrich Cat# V8137, RRID:AB_261889), anti-PKA Catalytic subunit (Abcam Cat# ab26322, RRID:AB_2170049), HRP-conjugated anti-mouse (Cell Signaling Technology Cat# 7076, RRID:AB_330924) and HRP-conjugated anti-rabbit (Cell Signaling Technology Cat# 7074, RRID:AB_2099233) were used for detection.

    Techniques: Expressing

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: STK25 inhibits PKA signaling by phosphorylating PRKAR1A

    doi: 10.1016/j.celrep.2022.111203

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: For Western blotting antibodies include: HRP conjugated anti-GAPDH (Cell Signaling Technology Cat# 3683, RRID:AB_1642205), anti-STK25 (Abcam Cat# ab157188, RRID:AB_2725788), anti-Flag (Sigma-Aldrich Cat# F3165, RRID:AB_259529), anti-GM130 (Cell Signaling Technology Cat# 12480, RRID:AB_2797933), anti-PRKAR1A (Abcam Cat# ab139695, RRID:AB_2893184), anti-pS77 PRKAR1A (Abcam Cat#ab139682, RRID:AB_2904566), anti-pS83 PRKAR1A (Abcam Cat#ab154851, RRID:AB_2904567), anti-PRKAR2A (ProteinTech Cat# 10142-2-AP), anti-phospholamban (Cell Signaling Technology Cat# 14562, RRID:AB_2798511), anti-phospho-phospholamban -pS16/T17 (Cell Signaling Technology Cat# 8496, RRID:AB_10949102), anti-Ryanodine receptor 2 (Abcam Cat#ab196355, RRID:AB_2904568), anti-p-Ryanodine receptor 2-pS2808 (Abcam Cat# ab59225, RRID:AB_946327), anti-TnI (Cell Signaling Cat# 4002), anti-TnI-pS23/S24 (Cell Signaling Cat# 4004), anti-V5 (Sigma-Aldrich Cat# V8137, RRID:AB_261889), anti-PKA Catalytic subunit (Abcam Cat# ab26322, RRID:AB_2170049), HRP-conjugated anti-mouse (Cell Signaling Technology Cat# 7076, RRID:AB_330924) and HRP-conjugated anti-rabbit (Cell Signaling Technology Cat# 7074, RRID:AB_2099233) were used for detection.

    Techniques: Recombinant, SYBR Green Assay, In Vitro, Activity Assay, Viability Assay, Sequencing, esiRNA, CRISPR, Knock-Out, Plasmid Preparation, Software

    A) Immunoblot of PRKAR1A phosphorylation sites S77 and S83 in STK25 +/+ and STK25 -/- cardiomyocyte protein lysates. B) Immunoblots of phosphorylation sites S77 and S83 of PRKAR1A in STK25 -/- cardiomyocytes transfected with empty vector (EV), wild type STK25 and kinase dead K49R/T174A (loaded in duplicate). C) Forskolin (10μM) stimulated STK25 +/+ and STK25 -/- cardiomyocytes immunoblotted for phosphorylation of PRKAR1A and RYR2. D) Immunoprecipitation of Flag-STK25 expressed in HEK293T cells and immunoblotted for PRKAR1A and GM130 (positive control binding partner). E) Co-immunoprecipitation of PRKAR1A-V5 with STK25 and PRKACA in HEK293T cells treated with forskolin (10μM). F) In vitro kinase assay of purified STK25 and PRKAR1A, immunoblotted for phosphorylation of S83 of PRKAR1A.

    Journal: bioRxiv

    Article Title: STK25 inhibits PKA signaling by phosphorylating PRKAR1A

    doi: 10.1101/2022.01.27.478084

    Figure Lengend Snippet: A) Immunoblot of PRKAR1A phosphorylation sites S77 and S83 in STK25 +/+ and STK25 -/- cardiomyocyte protein lysates. B) Immunoblots of phosphorylation sites S77 and S83 of PRKAR1A in STK25 -/- cardiomyocytes transfected with empty vector (EV), wild type STK25 and kinase dead K49R/T174A (loaded in duplicate). C) Forskolin (10μM) stimulated STK25 +/+ and STK25 -/- cardiomyocytes immunoblotted for phosphorylation of PRKAR1A and RYR2. D) Immunoprecipitation of Flag-STK25 expressed in HEK293T cells and immunoblotted for PRKAR1A and GM130 (positive control binding partner). E) Co-immunoprecipitation of PRKAR1A-V5 with STK25 and PRKACA in HEK293T cells treated with forskolin (10μM). F) In vitro kinase assay of purified STK25 and PRKAR1A, immunoblotted for phosphorylation of S83 of PRKAR1A.

    Article Snippet: For Western blotting antibodies include: HRP conjugated anti-GAPDH (Cell Signaling Technology Cat# 3683, RRID:AB_1642205), anti-STK25 (Abcam Cat# ab157188, RRID:AB_2725788), anti-Flag (Sigma-Aldrich Cat# F3165, RRID:AB_259529), anti-GM130 (Cell Signaling Technology Cat# 12480, RRID:AB_2797933), anti-PRKAR1A (Abcam Cat# ab139695, RRID:AB_2893184), anti-pS77 PRKAR1A (Abcam Cat#ab139682, RRID:AB_2904566), anti-pS83 PRKAR1A (Abcam Cat#ab154851, RRID:AB_2904567), anti-phospholamban (Cell Signaling Technology Cat# 14562, RRID:AB_2798511), anti-phospho-phospholamban -S16/T17 (Cell Signaling Technology Cat# 8496, RRID:AB_10949102), anti-Ryanodine receptor 2 (Abcam Cat#ab196355, RRID:AB_2904568), anti-p-Ryanodine receptor 2-S2808 (Abcam Cat# ab59225, RRID:AB_946327), anti-V5 (Sigma-Aldrich Cat# V8137, RRID:AB_261889), anti-PKA Catalytic subunit (Abcam Cat# ab26322, RRID:AB_2170049), HRP-conjugated anti-mouse (Cell Signaling Technology Cat# 7076, RRID:AB_330924) and HRP-conjugated anti-rabbit (Cell Signaling Technology Cat# 7074, RRID:AB_2099233) were used for detection.

    Techniques: Western Blot, Transfection, Plasmid Preparation, Immunoprecipitation, Positive Control, Binding Assay, In Vitro, Kinase Assay, Purification

    Fgf21 protected against structural and electrical remodeling in the atria. (A) Immunofluorescence analysis was performed to analyze myosin heavy chain (MHC). (B) Summary of the relative fluorescence intensity of MHC in each group of HL-1 atrial cells. (C) The expression of Troponin I, CACNA1C, Calpain, p-RyR2, and t-RyR2 at protein levels in each group of HL-1 atrial cells. * p < 0.05.

    Journal: Frontiers in Cardiovascular Medicine

    Article Title: Fibroblast Growth Factor 21 Protects Against Atrial Remodeling via Reducing Oxidative Stress

    doi: 10.3389/fcvm.2021.720581

    Figure Lengend Snippet: Fgf21 protected against structural and electrical remodeling in the atria. (A) Immunofluorescence analysis was performed to analyze myosin heavy chain (MHC). (B) Summary of the relative fluorescence intensity of MHC in each group of HL-1 atrial cells. (C) The expression of Troponin I, CACNA1C, Calpain, p-RyR2, and t-RyR2 at protein levels in each group of HL-1 atrial cells. * p < 0.05.

    Article Snippet: After transferring onto a PVDF membrane, protein was incubated with primary antibodies against Fgf21, Sirt1, Troponin I, t-CamKII, Calpian 1, NOX2, NOX4, GAPDH (Abcam), Nrf2, MHC, SOD2, UCP3, p-RyR2, β-Klotho (Abclonal), ox-CamKII (GeneTex), RyR2, FgfR1 (Proteintech), L-type calcium channel α1C-subunit (LCC) (Affinity), α-SMA, Collagen-1A1, CTGF, and Tubulin (Servicebio).

    Techniques: Immunofluorescence, Fluorescence, Expressing

    Nuclear factor erythroid 2-related factor 2 (Nrf2) positively regulated anti-oxidative genes expression induced by Fgf21. (A) Western blot was conducted to evaluate the expression of SOD2 and UCP3 in HL-1 atrial cells by knockdown of Nrf2 with si-Nrf2. (B) Western blot was conducted to evaluate the expression of Troponin I, Tgf-β, ox-CaMKII, t-CaMKII, CACNA1C, Calpain, p-RyR2, and t-RyR2 in HL-1 atrial cells by knockdown of Nrf2 with si-Nrf2. * p < 0.05.

    Journal: Frontiers in Cardiovascular Medicine

    Article Title: Fibroblast Growth Factor 21 Protects Against Atrial Remodeling via Reducing Oxidative Stress

    doi: 10.3389/fcvm.2021.720581

    Figure Lengend Snippet: Nuclear factor erythroid 2-related factor 2 (Nrf2) positively regulated anti-oxidative genes expression induced by Fgf21. (A) Western blot was conducted to evaluate the expression of SOD2 and UCP3 in HL-1 atrial cells by knockdown of Nrf2 with si-Nrf2. (B) Western blot was conducted to evaluate the expression of Troponin I, Tgf-β, ox-CaMKII, t-CaMKII, CACNA1C, Calpain, p-RyR2, and t-RyR2 in HL-1 atrial cells by knockdown of Nrf2 with si-Nrf2. * p < 0.05.

    Article Snippet: After transferring onto a PVDF membrane, protein was incubated with primary antibodies against Fgf21, Sirt1, Troponin I, t-CamKII, Calpian 1, NOX2, NOX4, GAPDH (Abcam), Nrf2, MHC, SOD2, UCP3, p-RyR2, β-Klotho (Abclonal), ox-CamKII (GeneTex), RyR2, FgfR1 (Proteintech), L-type calcium channel α1C-subunit (LCC) (Affinity), α-SMA, Collagen-1A1, CTGF, and Tubulin (Servicebio).

    Techniques: Expressing, Western Blot