Journal: Journal of Cardiovascular Pharmacology
Article Title: Transient Receptor Potential Vanilloid Type 1 Protects Against Pressure Overload–Induced Cardiac Hypertrophy by Promoting Mitochondria-Associated Endoplasmic Reticulum Membranes
doi: 10.1097/FJC.0000000000001301
Figure Lengend Snippet: TRPV1 activation protected against PE-induced cardiomyocyte hypertrophy in vitro. A, Representative images and analysis of the cell surface area of neonatal rat cardiomyocytes stained with cTnT (green) and DAPI (blue) (n = 5, scale bar = 10 μm). B, The ANP, BNP, β-MHC protein expression in primary cardiomyocytes treated with capsaicin and PE (n = 5). C, Quantitative polymerase chain reaction analyses of the mRNA levels of ANP, BNP, β-MHC in the indicated groups. n = 5 per group. * P < 0.05 compared with control, # P < 0.05 compared with PE group.
Article Snippet: After the nonspecific binding sites were blocked in Tris-buffered saline containing 5% nonfat dry milk for 1 hour, the membranes were incubated at 4°C overnight with primary antibodies, including mouse anti-TRPV1 (1:1000; cat. No. sc-398417; Santa Cruz Biotechnology, Santa Cruz, CA), rabbit anti–voltage-dependent anion channels (1:1000; cat. No. 10866-1-AP; Proteintech, Wuhan, China), rabbit anti-inositol 1,4,5-trisphosphate receptor (IP3R) (1:1000; cat. No. 19962-1-AP; Proteintech), rabbit anti-P-AMPK (1:1000; cat. No. 8208; Cell Signaling Technology, Danvers, MA), rabbit anti-AMPK (1:1000; cat. No. 5832; Cell Signaling Technology), rabbit anti-MFN2 (1:1000; cat. No. 12186-1-AP; Proteintech), rabbit anti-atrial natriuretic peptide (ANP) (1:500; cat. No. 27426-1-AP; Proteintech), rabbit anti-B-type natriuretic peptide (BNP) (1:500; cat. No. 13299-1-AP; Proteintech), and rabbit anti-beta-myosin heavy chain (β-MHC) (1:500; cat. No. 10799-1-AP; Proteintech).
Techniques: Activation Assay, In Vitro, Staining, Expressing, Real-time Polymerase Chain Reaction
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