Modulation of Ca2+-induced Ca2+ release by ubiquitin protein ligase E3 component n-recognin UBR3 and 6 in cardiac myocytes

Figure 1.. Ca2+ transient amplitude in cardiomyocytes.

Figure 2.. Protein expression of the Ca2+-handling proteins on the sarcolemmal and sarcoplasmic reticulum membranes in UBR3/6 knockdown NRVMs.

Figure 3.. Effects of UBR3/6 knockdown on Cav1.2 channel currents.

Figure 4.. Cycloheximide blocking assay of Cav1.2 channels in UBR3/6 knockdown NRVMs. (A) Effect of cycloheximide (CHX) on Cav1.2 channel expression in UBR3 knockdown NRVMs. NRVMs were transfected with or without UBR3 siRNA for 24 h, then treated with CHX (100 µg/ml). Aliquots of the cells were collected at 0, 4, 8, and 12 h after CHX treatment for western blot analysis. A typical example of a western blot analysis (left panel) and the summarized data (right panel) are shown. GAPDH served as a loading control (n = 3, *p < 0.01). Following the treatment of CHX, the Cav1.2 protein level showed a time-dependent decrease. The increase in Cav1.2 protein in UBR3 knockdown cells was not counteracted. (B) Effect of CHX on Cav1.2 channel expression in UBR6 knockdown NRVMs. NRVMs were transfected with or without UBR6 siRNA for 24 h then treated with CHX (100 µg/ml). Aliquots of the cells were collected at 0, 4, 8, and 12 h after CHX treatment for western blot analysis. A typical example of a western blot analysis (left panel) and the summarized data (right panel) are provided. GAPDH served as a loading control (n = 3, *p < 0.01). Following the treatment of CHX, the Cav1.2 protein level showed a…