Fig 2: Loss of NEK1 during meiosis is associated with hyper-phosphorylation and reduced ADD1 protein in a PP1?-dependent manner.(a) Quantitation of ADD1 protein levels in Nek1+/+ testis extracts relative to GAPDH control. (b) ADD1 and MYO10 protein levels, expressed as a ratio of Nek1kat2j/kat2j / Nek1+/+, as determined by mass spectrometry. (c) ADD1 phosphopeptide and deaminated peptide sequence determined by mass spectrometry to be higher in Nek1kat2j/kat2j testis extracts. (d) Quantitation by western blotting of ADD1 protein levels in oocytes at 0h and after 6h of culture. (e) Quantification of ADD1 protein levels in wildtype oocytes cultured for 6h in vehicle (ethanol) and the PP1 inhibitor Calyculin A (CLA) (at doses of 2 and 20 nM). All values are means ± Standard deviation. * Indicate statistically significant differences (Unpaired t test p < 0.05 and one-way ANOVA followed by Dunnett’s multiple comparisons test, p < 0.05, for cultures with inhibitor)