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

Journal: Reproductive Biology and Endocrinology : RB&E

Article Title: Expression of insulin-like 3 (INSL3) and differential splicing of its receptor in the ovary of rhesus macaques

doi: 10.1186/1477-7827-8-150

Figure Lengend Snippet: INSL3 protein expression in selected macaque tissues . a . Western blot analysis of INSL3 in selected endocrine-reproductive tissues. Eighty μg of total protein isolated from the ovary (Ov), testis (Te), hypothalamus (Hyp) and pituitary (Pit) was loaded in each lane. β-ACTIN (~47 kDa) was used as an internal loading control, and PageRuler™ Plus Prestained Protein Ladder (Fermentas) was used as molecular weight marker (MW). b . Immunohistochemistry (IHC) detection of INSL3 protein in the macaque ovary. Positive signal (brown) of INSL3 was localized in the thecal cells (The) surroundingantral follicles (AF), but not in other cell types within the ovary (A, B). Normal rabbit serum stained ovary section was used as negative control (C). GC, granulosa cells. c . INSL3 levels in monkey sera and follicular fluid during controlled ovarian stimulation (COS) protocols at 0-h and 36-h post hCG treatment. F serum follicular, female serum at follicular phase; F serum luteal, female serum at luteal phase; M, male. The value is presented as mean ± SE. Tissue lysate, sections, blood and follicular fluid were isolated from 3-4 different animals.

Article Snippet: The size of the detected protein band was determined by the PageRuler™ Plus Prestained Protein Ladder (Fermantas International Inc., Burlington, Ontario, Canada) separated on the same gel.

Techniques: Expressing, Western Blot, Isolation, Molecular Weight, Marker, Immunohistochemistry, Staining, Negative Control