Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Nanoscale coupling of junctophilin-2 and ryanodine receptors regulates vascular smooth muscle cell contractility

doi: 10.1073/pnas.1911304116

Figure Lengend Snippet: Jph2 knockdown has no effect on Ca 2+ spark frequency, amplitude, or kinetics. ( A ) Representative confocal Ca 2+ images of pressurized (60 mmHg), Fluo-4-AM–loaded cerebral arteries treated with control or Jph2 -targeting morpholinos. Colored boxes show selected ROIs where Ca 2+ sparks occurred. (Scale bar, 10 µm.) ( B ) Representative changes in fractional fluorescence ( F / F 0 ) as a function of time for ROIs in A . The trace color corresponds to the color of the respective ROI box. ( C ) Summary data showing the Ca 2+ spark frequency (in Hertz) normalized to surface area (in Hertz per 100 square micrometers) in cerebral arteries ( n = 5 to 6 cerebral arteries/group from 4 animals), as well as the amplitude ( F / F 0 ), half-duration [half-time ( t 1/2 ), in seconds], rise time ( t 1/2 , in seconds), and decay time ( t 1/2 , in seconds) of individual Ca 2+ spark events recorded from each group ( n = 616 events for control, n = 601 events for Jph2 -targeted). There were no significant differences.

Article Snippet: Rows were successively loaded with Wes antibody diluent blocking buffer; anti-JPH2 (ab116077; Abcam) or anti–β-actin (ab8227, Abcam) primary antibodies, diluted 1:20 and 1:1,500, respectively; horseradish peroxidase (HRP)-conjugated anti-rabbit secondary antibody (1×; ProteinSimple); and a luminol–peroxide mix.

Techniques: Fluorescence