Journal: bioRxiv

Article Title: A complex phenotypic assay of mammalian oocyte maturation identifies compounds that block meiotic progression for non-hormonal contraceptive discovery

doi: 10.1101/2025.10.15.682568

Figure Lengend Snippet: Compounds from a bioactives library (Selleckchem Bioactives Library II) were filtered to remove those with undesirable chemical properties and prioritize a subset for testing within the phenotypic screening assay. Hits identified from the initial counterscreen assay then underwent hit validation, including confirmation using an independent source, concentration-response, and prolonged culture treatment. For GV-arresting compounds, an additional PDE3A activity counterscreen assay was conducted as well as further compound profiling, including chemical validation using structural analogs and reversibility studies.

Article Snippet: Thus, in the Selleckchem Bioactives Compound Library II used in this study, we filtered and removed PAINS and cytotoxic compounds to prevent detection of undesired, non-selective hits during screening ( ).

Techniques: Screening Assay, Biomarker Discovery, Concentration Assay, Activity Assay

Journal: bioRxiv

Article Title: A complex phenotypic assay of mammalian oocyte maturation identifies compounds that block meiotic progression for non-hormonal contraceptive discovery

doi: 10.1101/2025.10.15.682568

Figure Lengend Snippet: ( A ) Structural similarity chart of compounds from the entire Selleckchem Bioactives Library II (n=5307). Red dots indicate compounds deprioritized due to unprogressable (BAD-SMARTS) or pan-assay interference (PAINS) features. ( B ) Structural similarity chart of high-priority compounds filtered from the Selleckchem Bioactives Library (n=2812). Purple dots indicate the final compounds selected for the phenotypic oocyte maturation screening assay. ( C ) Overview of phenotypic oocyte maturation screening assay. Denuded oocytes at the GV stage were placed in an overnight incubator for in vitro maturation (IVM) with either DMSO (negative control), 10 µM Milrinone (positive control), or 10 µM compound-of-interest. Brightfield images were taken before and after IVM to determine oocyte survival and maturation status. Asterisk indicates germinal vesicle and black arrow indicates the extruded polar body. ( D ) Representative images of oocytes treated with DMSO (negative control) and 10 µM Milrinone (positive control) before and after IVM. ( E ) Graph of MII incidence for negative and positive controls across phenotypic oocyte screening assay plates and of Z’-factor to determine assay quality. GV, germinal vesicle; GVBD, germinal vesicle breakdown; MII, meiosis II.

Article Snippet: Thus, in the Selleckchem Bioactives Compound Library II used in this study, we filtered and removed PAINS and cytotoxic compounds to prevent detection of undesired, non-selective hits during screening ( ).

Techniques: Pan Assay, Screening Assay, In Vitro, Negative Control, Positive Control