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. Author manuscript; available in PMC: 2019 Nov 13.
Published in final edited form as: J Cell Physiol. 2018 Mar 25;233(9):7239–7252. doi: 10.1002/jcp.26557

FIGURE 5.

FIGURE 5

Involvement of PP1 in manifestation of cAMP/PKA axis for regulation of GSK3 function. (a) Comparison of phosphatase activity in soluble protein fraction of the sAC knockout to wild-type mice sperm cells; PP1 activity was determined using both I2 and okadaic acid (2 nM), separately. Data represent mean ± SEM of n = 5 for all the values. **p ≤ 0.01 vs sAC+/+ values. (b) In vitro regulation of Ser 21/9-phosphorylation profile of GSK3 by PP1 under influence of PKA in the sAC knockout mice sperm. (c) Effect of PKA inhibition in phosphatase activity in WT mouse sperm extract. (d) In vitro regulation of phospho-Ser-GSK3 profile by PP1 under influence of PKA in WT sperm. Pre-treatment using H89 was done for 30 min for the designated sets followed by incubation with calyculin A for another 30 min at 37 °C. (e) Pull down assay to check interactions among GSK3, PP1 and PKA using mouse rPP1γ2. All the probing were done using corresponding rabbit raised antibodies except for GSK3 where anti-mouse antibody was used