bk 036s cdc42  (Cytoskeleton Inc)

Journal: Diabetes, Metabolic Syndrome and Obesity

Article Title: Empagliflozin Induces Vascular Relaxation in Rat Coronary Artery Due to Activation of BK Channels

doi: 10.2147/DMSO.S419125

Figure Lengend Snippet: Empagliflozin induces vasodilation in KCl-constricted coronary arteries.

Article Snippet: The PVDF membranes were incubated with specific primary antibodies, with β-actin (ab20272, Abcam, 1:5000), BK-α (APC-021, Alomone Labs, 1:500), BK-β1 (APC-036, Alomone Labs, 1:1000), Sirt1 (ab110304, Abcam, 1:2000) and Nrf2 (SAB5700720, Sigma, 1:1000).

Techniques:

Journal: Frontiers in Cardiovascular Medicine

Article Title: Large-conductance Ca 2 + -activated K + channel β1-subunit maintains the contractile phenotype of vascular smooth muscle cells

doi: 10.3389/fcvm.2022.1062695

Figure Lengend Snippet: BK Ca -β1 expression is reduced in the carotid arteries of rat balloon-injured model. Representative images of H&E (A) and immunohistochemical staining of BK Ca −α and BK ca −β1 (B) in injured and sham-operated carotid arteries of rat balloon-injured model at 14 days post injury. Scale bar = 100 μm. (C) Quantified mRNA levels of BK ca −α and BK ca −β1 analyzed by qRT-PCR. Western blot images (D) and quantification (E) for indicated proteins. Eukaryotic initiation factor 5 (eIF5) was used as an internal control. Data are presented as means ± SEM ( n = 4∼6 for each group, ns, not significant, * P < 0.05, ** P < 0.01 vs. sham).

Article Snippet: An antibody against BK Ca -β1 (APC-036) used for immunohistochemical staining and western blot was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Expressing, Immunohistochemical staining, Staining, Quantitative RT-PCR, Western Blot, Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: Large-conductance Ca 2 + -activated K + channel β1-subunit maintains the contractile phenotype of vascular smooth muscle cells

doi: 10.3389/fcvm.2022.1062695

Figure Lengend Snippet: BK Ca -β1 expression is positively associated with VSMC differentiation markers in vitro . Western blot images (A) and quantification (B) of BK ca −α, BK ca −β1, and VSMC differentiation markers expression in VSMCs of passage 1 and passage 6. Western blot images (C) and quantification (D) of BK ca −α, BK ca −β1, and VSMC differentiation markers expression in VSMCs after PDGF-BB treatment (25 μg/L). Western blot images (E) and quantification (F) of BK ca −α, BK ca −β1, and VSMC differentiation markers expression in VSMCs treated with TGF-β (2.5 μg/L). VSMCs were subjected to serum-starvation for 24 h and then treated with the indicated stimulation for 48 h. Passages 3-5 of VSMCs were used in (C) through (F) . Eukaryotic initiation factor 5 (eIF5) was used as an internal control. Data are presented as means ± SEM ( n = 5 for each group, ns, not significant, * P < 0.05, ** P < 0.01, and *** P < 0.001 vs. control).

Article Snippet: An antibody against BK Ca -β1 (APC-036) used for immunohistochemical staining and western blot was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Expressing, In Vitro, Western Blot, Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: Large-conductance Ca 2 + -activated K + channel β1-subunit maintains the contractile phenotype of vascular smooth muscle cells

doi: 10.3389/fcvm.2022.1062695

Figure Lengend Snippet: Decreased BK ca −β1 expression is mediated by elevated PDGF-BB in vivo . (A) Quantified mRNA levels of PDGF-B and TGF-β analyzed by qRT-PCR in injured and sham-operated carotid arteries. Western blot images (B) and quantification (C) of BK ca −α and BK ca −β1 in the carotid arteries of mice injected with saline or PDGF-BB via tail vein. Data are presented as means ± SEM ( n = 6 for each group, ns, not significant, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 vs. control).

Article Snippet: An antibody against BK Ca -β1 (APC-036) used for immunohistochemical staining and western blot was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Expressing, In Vivo, Quantitative RT-PCR, Western Blot, Injection, Saline, Control

Journal: Frontiers in Cardiovascular Medicine

Article Title: Large-conductance Ca 2 + -activated K + channel β1-subunit maintains the contractile phenotype of vascular smooth muscle cells

doi: 10.3389/fcvm.2022.1062695

Figure Lengend Snippet: BK Ca -β1 is essential for VSMCs to maintain a differentiated phenotype. (A) Representative micrographs of VSMCs treated with BK ca −β1 siRNA or scramble siRNA transfection. (B) Representative immunofluorescent images of F-actin of VSMCs stained with phalloidin (red). Nuclei were stained with DAPI (blue). Scale bar = 25 μm. (C) The percentage of spindle-like or polygonal-like cells in each group. Western blot images (D) and quantification (E) of BK ca −β1, α-SMA, and SM22α. Representative images (F) and quantification (G) of the areas of collagen gel in each group. The size of the collagen gel in each well was calculated compared to the total area of the well. Data are presented as means ± SEM ( n = 5∼6 for each group, *** P < 0.001 vs. scramble).

Article Snippet: An antibody against BK Ca -β1 (APC-036) used for immunohistochemical staining and western blot was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Transfection, Staining, Western Blot

Journal: Frontiers in Cardiovascular Medicine

Article Title: Large-conductance Ca 2 + -activated K + channel β1-subunit maintains the contractile phenotype of vascular smooth muscle cells

doi: 10.3389/fcvm.2022.1062695

Figure Lengend Snippet: BK Ca -β1 knockdown induces proliferative, migratory, and synthetic phenotype of VSMCs. (A) Proliferation curves of VSMCs transfected with BK ca −β1 or scrambled siRNA. Representative images (B) and quantification (C) of migration assay. (D) Quantified mRNA levels of inflammatory factors analyzed by qRT-PCR. Representative images (E) and quantification (F) of activities of MMP-9 and MMP-2 tested using gelatin zymography. Data are presented as means ± SEM ( n = 5∼6 for each group, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001 vs. scramble).

Article Snippet: An antibody against BK Ca -β1 (APC-036) used for immunohistochemical staining and western blot was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Knockdown, Transfection, Migration, Quantitative RT-PCR, Zymography

Journal: Frontiers in Cardiovascular Medicine

Article Title: Large-conductance Ca 2 + -activated K + channel β1-subunit maintains the contractile phenotype of vascular smooth muscle cells

doi: 10.3389/fcvm.2022.1062695

Figure Lengend Snippet: Decreased BK ca −β1 expression is correlated with VSMC dedifferentiation and atherosclerosis in human. Western blot images (A) and quantification (B) of BK ca −α and BK ca −β1 expression in human primary aortic VSMCs after PDGF-BB treatment (25 μg/L). Western blot images (C) and quantification (D) of BK ca −α and BK ca −β1 expression in human primary aortic VSMCs treated with TGF-β (2.5 μg/L). VSMCs were subjected to serum-starvation for 24 h and then treated with the indicated stimulation for 48 h. Passages 3–5 of VSMCs were used. (E) Representative western blot images of the protein levels of BK ca −α and BK ca −β1 in lysates of human carotid endarterectomy artery (CEA) and control internal mammary arteries (IMA). (F) Quantification was performed by calculating the ratio between the levels of BK ca −β1 and BK ca −α. Eukaryotic initiation factor 5 (eIF5) was used as an internal control. Data are presented as means ± SEM (n = 3∼5 for each group, ns, not significant, * P < 0.05, ** P < 0.01 vs. control).

Article Snippet: An antibody against BK Ca -β1 (APC-036) used for immunohistochemical staining and western blot was purchased from Alomone labs (Jerusalem, Israel).

Techniques: Expressing, Western Blot, Control