Figure 4

Subcellular location of ryanodine receptor, thapsigargin-sensitive Ca²⁺-ATPase, and endomembranes in mouse granulosa cells. The first block shows the intracellular location of the ryanodine receptor visualized by BODIPY FL-ryanodine. The second block depicts the intracellular location of thapsigargin-sensitive Ca²⁺-ATPase using BODIPY-FL thapsigargin. The third block shows the distribution of endomembranes by means of the BODIPY-FL conjugate of brefeldin A. All fluorescent probes were assayed at 1 μM (Yellow color). The second column shows the signal of the nuclear marker DAPI (Cyan color), whereas the third column depicts the merging the first and second columns. The fourth column is a magnification of a selected sector of the merge image. The results strongly suggest the intranuclear presence of all the Ca²⁺-handling proteins and the endomembranes in the mouse granulosa cells. Scale bar corresponds to 20 μm. Representative of 5 independent experiments.