Figure 6.

JP2 mediates the molecular coupling of RyR and Cav1 in MASMCs. A, Western blotting analyses were performed using the protein lysate (30 μg) from mouse mesenteric artery tissues treated with siRNA for 4 days. The band density of JP2 was normalized to that of α-SM actin (JP2/α-SM actin). The ratio of siJP2 was normalized to that of siControl. B, summarized effects of siControl and siJP2 on JP2 protein expression in the mesenteric artery. C, representative transmitted light and immunofluorescent images of mMASMCs treated with siControl or siJP2. Myocytes were labeled with an anti-JP2 antibody and observed using a confocal microscope. D, summarized effects of siControl (n = 23) and siJP2 (n = 14) on JP2 expression estimated by fluorescent intensity. E and F, RyR and Cav1 in mMASMCs treated with siControl (E) or siJP2 (F) were labeled with the specific antibody and observed under a TIRF microscope. Fluorescent signals corresponding to Cav1, RyR, and their co-localization are colored in red, green, and yellow (denoted by arrowheads in merged), respectively. The yellow puncta in co-localized indicate only the overlapping signals of Cav1 and RyR. G and H, the co-localization ratio of RyR (G) or Cav1 (H) particles in myocytes treated with siControl (n = 9) or siJP2 (n = 8). *, p < 0.05; **, p < 0.01; the Student's t test. Scale bars indicate 10 μm (C, E, and F).