5 sodium channel blocker  (Santa Cruz Biotechnology)

Journal: Biomedicines

Article Title: Prior Aerobic Exercise Training Fails to Confer Cardioprotection Under Varying Exercise Volumes in Early Post-Infarction Cardiac Remodeling in Female Rats

doi: 10.3390/biomedicines13092221

Figure Lengend Snippet: Calcium-handling proteins. ( A ) PLB, total phospholamban; ( B ) PLB(Thr17/Ser16), phospholamban phosphorylated at serine 16 and threonine 17; ( C ) NCX1, sodium/calcium exchanger 1; ( D ) SERCA2a, sarco/endoplasmic reticulum Ca 2+ -ATPase; ( E ) LTCC, L-type Ca v 1.2 voltage-gated calcium channel; ( F ) RyR, ryanodine receptor. GAPDH: glyceraldehyde 3-phosphate dehydrogenase constitutive protein. Boxplots (black squares with whiskers) with individual data points (small circles of different colors) and means (red circles) or 20% trimmed means (red squares). The p -value displayed above the boxplots refers to the ANOVA. Brackets were added to the graphs to represent significant differences in pairwise comparisons. ( B – D , F ) charts: Fisher’s one-way ANOVA and pairwise comparisons via Student’s t test. ( A , E ) charts: Welch’s ANOVA with trimmed means and pairwise comparisons via Yuen’s trimmed test. The GAPDH-LTCC and GAPDH-RyR are equivalent because the same membrane was used to detect LTCC and RyR through membrane stripping for Western blotting.

Article Snippet: The transferred proteins were then incubated at ~4 °C (overnight) with the following primary rabbit antibodies: anti-L-type high-voltage calcium channel Ca v 1.2 (LTCC) (1:10,000 ACC-003, Alomone, Jerusalem, Israel), anti-sarco/endoplasmic reticulum Ca 2+ -ATPase (SERCA2a) (1:1000 A010-20, Badrilla, Leeds, UK), anti-phospholamban phosphorylated at threonine 17 and serine 16 (PLB(Thr17/Ser16)) (1:5000 ab62170, Abcam, Cambridge, MA, USA), anti-sodium/calcium exchanger 1 (NCX1) (1:5000 ab177952, Abcam, Cambridge, MA, USA), anti-4-hydroxynonenal (4-HNE) (1:4000 ab46545, Abcam, Cambridge, MA, USA), and anti-GAPDH (1:20,000 D16H11, Cell Signaling Technology, Danvers, MA, USA), as well as the following primary mouse antibodies: anti-ryanodine receptor (RyR) (1:2000 ab2868, Abcam, Cambridge, MA, USA), and anti-total phospholamban.

Techniques: Membrane, Stripping Membranes, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Chronic Partial Sleep Deprivation Increased the Incidence of Atrial Fibrillation by Promoting Pulmonary Vein and Atrial Arrhythmogenesis in a Rodent Model

doi: 10.3390/ijms25147619

Figure Lengend Snippet: Histological and Western blot analyses were performed on the atrial appendages of both left atriums (LAs) and right atriums (RAs) in both control and SD rats. ( A ) Masson staining of the atrial appendages in the LAs and RAs of both the control and SD groups highlighted a significant increase in fibrosis and collagen deposition in the SD LAs and RAs when compared to the corresponding structures in the control group. ( B ) Expression and phosphorylation levels of calcium-handling proteins, upstream kinases, and β1-adrenoceptor (β1-AR) in the control and SD LAs. ( C ) Expression and phosphorylation levels of calcium-handling proteins, upstream kinases, and β1-AR in the control and SD RAs. The expression level of G protein-coupled receptor kinase 2 (GRK2) was significantly reduced in the LAs of SD rats in comparison to those of control rats. β1-AR, beta-1 adrenoceptor; CaMKII, Ca 2+ /calmodulin-dependent kinase II-δ; pCaMKII, phosphorylated CaMKII at Thr286; NCX, Na + /Ca 2+ exchanger; Cav1.2, Cav1.2 L-type calcium channel; RyR2, ryanodine receptor type 2; pRyR S2808, phosphorylated RyR2 at Ser2808.

Article Snippet: All blots were probed with primary antibodies against a beta-1 adrenergic receptor (β1-AR, #PA1-049, Thermo Fisher Scientific, Waltham, MA, USA), G protein-coupled receptor kinase 2 (GRK2, #SC-562, Santa Cruz Biotechnology, Dallas, TX, USA), Ca 2+ /calmodulin-dependent kinase II-δ (CaMKII-δ, #GTX111401, GeneTex, Irvine, CA, USA), phosphorylated CaMKII at Thr 286 (pCaMKII, #ab32678, Abcam, Cambridge, UK), ryanodine receptor type 2 (RyR2, #MA3-916, Thermo Fisher Scientific, Waltham, MA, USA), phosphorylated RyR2 at Ser 2808 (pRyR S2808, #A010-30AP, Badrilla, Leeds, UK), catalytic subunit of protein kinase A (PKAc, #610981, BD Transduction Laboratories, San Jose, CA, USA), Na + /Ca 2+ exchanger (NCX, #R3F1, Swant, Burgdorf, Switzerland), Cav1.2 L-type calcium channel (Cav1.2, #ACC-003, Alomone Lab, Jerusalem, Israel), and glyceraldehyde-3-phosphate dehydrogenase (GADPH, #M171-7, MBL, Nagoya, Japan).

Techniques: Western Blot, Control, Staining, Expressing, Comparison

Journal: bioRxiv

Article Title: Exploring the functional, protective, and transcriptomic effects of GIP on cytokine-exposed human pancreatic islets and EndoC-βH5 cells

doi: 10.1101/2024.06.17.599254

Figure Lengend Snippet: (a) Secreted insulin during low glucose (2 mM, left) and during high glucose (20 mM, right) with increasing concentrations (0.025-1000 nM) of GIP. Angled lines on the x-axis denote the transition from x10 increments in GIP dose to 1000 nM GIP. Data are represented as mean ± SD of 4-6 independent experiments. Asterisks (*) indicate statistical significance using mixed-effect analysis/model with Dunnett’s multiple comparisons test each GIP dose compared to 0 nM at 2 or 20 mM glucose, respectively, **p < 0.01, ***p < 0.001. Secreted insulin during low glucose (2 mM, left) and during high glucose (20 mM, right) upon treatment (10 μM each) with H-89 (PKAi), bisindolylmaleimide I (PKCi), nimodipine (LTCCi), KN-93 (CaMK2i), diazoxide (DIA) or vehicle control (0.1% DMSO) without (grey bars) or with GIP (2.5 nM, green bars). Data are represented as mean ± SD of 4 independent experiments. Asterisks (*) indicate statistical significance using paired RM two-way ANOVA with Dunnett’s multiple comparisons test comparing inhibitors to CTRL or GIP vehicle control (DMSO) at 2 or 20 mM glucose, respectively, * p <0.05, ** p < 0.01, *** p <0.001.

Article Snippet: The acute effect of GIP on GSIS was also assessed in the presence of a selection of intracellular signalling inhibitors; H89 (a protein kinase A (PKA) inhibitor, Tocris Bioscience, UK), Bisindolylmaleimide I (a protein kinase C (PKC) inhibitor, Sigma-Aldrich, USA), Nimodipine (an L-type calcium channel (LTCC) inhibitor, Sigma-Aldrich, USA), KN-93 (a calcium/calmodulin-dependent protein kinase II (CaMK2) inhibitor, Sigma-Aldrich, USA), Diazoxide (an inhibitor of K ATP -channel closure and membrane depolarisation, Sigma-Aldrich, USA) or vehicle control (equivalent dilution of 0.1% DMSO).

Techniques: