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Fig. 11. Imaging of exocytotic discharge of renin and neutral red by ML-7. Cells on postconfluent day 2 were plated on cover slips were permeabilized and stained with antibody to renin. Many cytoplasmic stained granules are seen (A, control). When cells were pretreated with ML-7 (6 × 10−5 M) for 2 min and stained with renin antibody, no granules were observed (ML-7). Scale bar: 10 μm. Cells were incubated with 100 μM neutral red for 3 h, washed with fresh DMEM, and photographed with a light microscope. Numerous neutral red stained pink granules are seen (B). N denotes nucleus. Scale bar: 3.33 μm. (C, D) When neutral red-loaded granules were viewed at the isosbestic absorbent light at 470 nm, the granules observed were black. White granules are likely discharged granules prior to imaging. When cells were perfused with ML-7 (6 × 10−5 M), #1, #2, and #3 neutral red-loaded granules were discharged over 200 msec. (D) The time-lapsed movie shows the discharge of neutral red-loaded secretory granules (Supplementary Video). DMEM, Dulbecco’s modified eagle medium.
Korean J Physiol Pharmacol 2022;26:479-499
© Korean J Physiol Pharmacol