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Fig. 3. Recovery from acidosis in isolated granulosa cells. Granulosa shells
were prepared by removing the oocyte from preantral follicles recovered from
10-day-old mice, and then loaded with SNARF-AM. (A) Left, a series of
three photomicrographs illustrating the procedure by which the oocyte
(arrowheads) is removed from a preantral follicle. A higher resolution
exemplar photomicrograph of the resulting granulosa shell is also shown
(center) and a fluorescence micrograph of SNARF-loaded shells (right). Scale
bar: 50 µm. (B) Recovery from acidosis in SNARF-loaded granulosa
shells following a NH4Cl pulse. Note the rapid increase in pH when
Na+ was returned to the bath (experiments were performed in
triplicate, n=23 in total). Note also that amiloride retarded
Na+-dependent acidosis correction (experiments were performed in
triplicate, n=23 in total). Amiloride was added at t=10
minutes. (C) Average rate of recovery from acidosis following
Na+ replacement. *, P<0.05.