Overview of the proteomics and phosphoproteomics experiments. (A) Outline of the phosphoproteomics experiment. Mos–MAPK-inhibited (20 µM U0126, purple) and control (DMSO, green) oocytes were collected at metaphase I in three biological replicates and processed for phosphoproteomics. For the complete dataset, see Data S1. (B) Unsupervised clustering of differentially phosphorylated peptides (rows) vs TMT channels (columns). Green: control, purple: Mos–MAPK inhibition. TMT reporter intensity values are scaled to a mean of 0 and SD of 1. (C) GO overrepresentation analysis of differentially phosphorylated proteins. The numbers indicate the gene set sizes and a number of differentially phosphorylated proteins. (D) Prediction of human effector kinase families from the differentially dephosphorylated sites. (E) Outline of the proteomics time course experiment. Mos–MAPK-inhibited (20 µM U0126, purple) and control (DMSO, green) oocytes were collected at the specified stages at the given times in two biological replicates. Samples were processed in three technical replicates for proteomics and analyzed by DIA. For the complete dataset, see Data S2. (F) Unsupervised clustering of centered DIA intensities of two biological and three technical replicates (columns). Green: control, purple: Mos–MAPK inhibition. (G) Log₁₀ normalized protein intensities for the indicated proteins. The mean and standard error of the two biological replicates and three technical repetitions of each are shown.