Journal: The Journal of physiology

Article Title: MitoBK Ca channel is functionally associated with its regulatory β1 subunit in cardiac mitochondria

doi: 10.1113/JP277769

Figure Lengend Snippet: A) Immunoprecipitation (IP) of BKCa alpha and BK-β1 subunit with an antibody against BKCa alpha (see methods). Western blots revealed the presence of GRIM 19 and SERCA as markers of mitochondrial and sarcoplasmic reticulum membranes (4A, lower panels). *The signal of BK-β1 in the whole lysate was detected with chemiluminescent (ECL) substrate Supersignal™ West Femto Maximun sensitivity (see methods). Taken together, these results indicates an association between mitoBKCa and its auxiliary β1 subunit. Having demonstrated this, we characterized the biophysical properties of mitoBKCa in Wt and β1-KO cardiac mitoplasts. B) Single-channel currents of mitoBKCa recorded from Wt mitoplasts. The Po of the channel was 0.9 ± 0.08, n=4, in average at +80 mV and the indicated matrix Ca2+. C) Representative traces of a mitoplast from the β1 KO, were no single-channel current was detected at the indicated voltages and matrix Ca2+ (upper traces). Few mitoplasts (5 out of 28, mitoplasts, n=5 hearts) showed mitoBKCa channel activity with a Po of 0.4 ± 0.004, n=3 at +40 mV in the indicated matrix Ca2+. D) Plots of I vs. V at 12 µM matrix Ca2+ from Wt and β1 KO mitoplasts (data from 6 and 3 patches, respectively). A slope conductance of 302±26 (n=4) and 335 ±15 pS (n=3) was calculated for Wt and β1 KO, respectively. E) Plots of Po vs. V for mitoBKCa channels, data points represent the average of 4 and 3 mitoplasts for Wt and BK-β1 KO, respectively. Bars SEM.

Article Snippet: Polyclonal antibody against BK Ca (Rabbit Anti-KCNMA1 Cat. APC-021 and APC-151), and polyclonal antibody for BK-β1 subunit (Rabbit Anti-KCNMB1, Cat. APC-036) were from Alomone labs.

Techniques: Immunoprecipitation, Western Blot, Activity Assay

Journal: Oncotarget

Article Title: Characterization of pressure-mediated vascular tone in resistance arteries from bile duct-ligated rats

doi: 10.18632/oncotarget.15409

Figure Lengend Snippet: A . Immunoblots for Ca v 1.2, TRPC3 and β-actin (loading control) from small mesenteric artery homogenates indicate no differences among both groups. The bar graphs show densitometry analyses for Ca v 1.2 and TRPC3. B . qPCR for Ca v 1.2. Representative IHC photographs and qPCR indicate that in the arteries from the BDL-rats, expressions of C ., D . BK Ca and E ., F . TRPC6 are reduced when compared to SHAM-rats ( n = 3-5/group). G . The immunoblots for both BK Ca and TRPC6 and their loading controls are shown. The bar graphs show densitometry analyses for BK Ca and TRPC6 ( n = 3/group). * P < 0.05, ** P < 0.01 when compared to SHAM-rats.

Article Snippet: Then, the sections were incubated with anti-TRPC6 or anti-BK Ca polyclonal antibodies (1:200, Alomone Labs, Israel) overnight at 4°C in a humidified chamber.

Techniques: Western Blot

Journal: American Journal of Physiology - Regulatory, Integrative and Comparative Physiology

Article Title: Long-term exposure to high-altitude chronic hypoxia during gestation induces neonatal pulmonary hypertension at sea level

doi: 10.1152/ajpregu.00123.2010

Figure Lengend Snippet: Components of the NO signaling pathway in LLL (open bars) and LHL (filled bars) lambs. Values are the means ± SE for the protein expression of eNOS ( A ), soluble guanylate cyclase (sGC, B ), phosphodiesterase 5 (PDE5, C ), and Ca 2+ -activated K + channels (BK Ca , D ). E : individual immunoblots. Significant differences ( P < 0.05) are LHL vs. LLL (†).

Article Snippet: Protein expression of eNOS, HO-1, sGC, BK Ca , PDE5, and β-actin was determined in total lung lysates by immunoblot with specific anti-eNOS monoclonal antibody (Transduction Laboratories), anti-HO-1 monoclonal antibody (Research Diagnostic), anti-sGC polyclonal antibody (Cayman Laboratories), anti-BK Ca polyclonal antibody (Alomone laboratories), anti-PDE5 monoclonal antibody (BD Transduction Laboratories), and anti-β-actin monoclonal antibody (Sigma) as described elsewhere ( ).

Techniques: Expressing, Western Blot