Journal: Cardiovascular Research

Article Title: Ryanodine receptor subtypes regulate Ca 2+ sparks/spontaneous transient outward currents and myogenic tone of uterine arteries in pregnancy

doi: 10.1093/cvr/cvaa089

Figure Lengend Snippet: Knockdown of RyR1/RyR2 reduced the interaction of BKCa channel α and β1 subunits in uterine arteries. (A) Representative immunoblots from five replicates show co-immunoprecipitation of BKCa channel α and β1 subunits in uterine arteries of non-pregnant (NUA) and pregnant (PUA) sheep. Uterine arteries were treated with scramble control siRNA or siRNAs for RyR1, RyR2, and RyR3, respectively, for 48 h. IgG was used as a control to show antibody specificity. (B) Representative confocal immunofluorescence images from five replicates show the co-localization of BKCa channel α and β1 subunits in uterine arteries of pregnant sheep after control siRNAs or RyR siRNAs treatments. The arteries were stained with antibodies against α (green) and β1 (red) subunits. Merged images show in yellow. The nuclear region was stained with DAPI and shows in blue. Scale bar: 100 µm. (C) Representative immunoblots from five replicates show co-immunoprecipitation of BKCa channel α and β1 subunits in uterine arteries of pregnant sheep after control siRNA or RyR siRNAs treatments. β-Actin blots showing equal total protein lysates (input).

Article Snippet: After blocking non-specific binding sites by dry milk, membranes were incubated with primary antibodies (1:1000 dilution) against rabbit polyclonal RyR1 (8153, Cell Signaling, Danvers, MA, USA), rabbit polyclonal RyR2 (ARR-002, Alomone Labs, Israel), rabbit polyclonal RyR3 (AB9082, EMD Millipore), mouse monoclonal BK Ca channel α, or mouse monoclonal BK Ca channel β1 (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Western Blot, Immunoprecipitation, Immunofluorescence, Staining

Journal: Cardiovascular Research

Article Title: Ryanodine receptor subtypes regulate Ca 2+ sparks/spontaneous transient outward currents and myogenic tone of uterine arteries in pregnancy

doi: 10.1093/cvr/cvaa089

Figure Lengend Snippet: Knockdown of RyR1/RyR2 downregulated the expression of BKCa channel β1 subunit in uterine arteries of pregnant animals. Uterine arteries of pregnant sheep were treated with scramble control siRNA or siRNAs for RyR1, RyR2, and RyR3, respectively. Protein abundance of BKCa channel α and β1 subunits was measured by western blot in uterine arteries 48 h after the treatment. (A) RyR1 siRNAs treatment. (B) RyR2 siRNAs treatment. (C) RyR3 siRNAs treatment. Data are means ± SEM from five animals of each group; independent-samples t-test; *P < 0.05, RyR siRNAs vs. control siRNA.

Article Snippet: After blocking non-specific binding sites by dry milk, membranes were incubated with primary antibodies (1:1000 dilution) against rabbit polyclonal RyR1 (8153, Cell Signaling, Danvers, MA, USA), rabbit polyclonal RyR2 (ARR-002, Alomone Labs, Israel), rabbit polyclonal RyR3 (AB9082, EMD Millipore), mouse monoclonal BK Ca channel α, or mouse monoclonal BK Ca channel β1 (Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Western Blot