FIGURE

Fig. 3

ID

ZDB-FIG-260207-3

Publication

Akhter et al., 2025 - KLHL13 functional defects cause neurodevelopmental disorder in humans that can be rescued via inhibition of AURKB in cellular and animal models

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Fig. 3

KLHL13 variants lead to increased expression and reduced ubiquitination levels of AURKB, resulting in abnormal cell-cycle regulation, which can be rescued by AZD1152-HQPA treatment. A. Immunoblot showing the expression of AURKB in KLHL13WT and variants overexpressing cells. B. Quantitative representation of AURKB in KLHL13WT and variants overexpressing cells revealed a significant increase in the AURKB levels in the variants (p.His60Arg, ∗∗P < .0117; p.Cys186Tyr, ∗P < .0193; p.Val594Glyfs∗6, ∗P < .0419) compared with KLHL13WT. C. Immunoprecipitation of AURKB in KLHL13WT and variants overexpressing cells, followed by Western blot analysis using ubiquitin and AURKB antibodies. The yellow dotted line box represents the area that was quantified for ubi-AURKB expression. D. Graph representing a significantly reduced AURKB ubiquitination level in KLHL13 variants (p.His60Arg, ∗∗P < .0037; p.Cys186Tyr, ∗∗P < .0046; p.Ala318Val, ∗P < .0118; p.Val594Glyfs∗6, ∗P < .0215) overexpressing cells compared with KLHL13WT. E. Graphical representation of KLHL13 constructs overexpressing cells at different cell-cycle phases. In contrast to untreated cells, AZD1152-HQPA (30 nM) treatment significantly (∗∗P < .001) improved mitosis and genomic stability in cells overexpressing variants harboring KLHL13 constructs. F. Confocal images of COS-7 cells at anaphase overexpressing KLHL13 NDD-associated variants, with or without AZD1152-HQPA (30 nM) treatment, and immunolabeled with anti-AURKB (orange), anti-GFP (green), and anti-β-tubulin (red) antibodies, whereas DNA content was marked with DAPI (blue). After treatment, AURKB localization at the central spindle was restored in cells expressing KLHL13 pathogenic variants. Scale bar: 20 μm. G. Quantitative representation of cells with AURKB localization at the central spindle vs chromosome arms. AZD1152-HQPA treatment significantly (∗∗ P < .001, ∗∗∗P < .0007) improved AURKB localization at the central spindles when compared with untreated cells. A total of 90 cells for each condition were imaged and plotted, with 30 cells quantified per experiment over 3 independent experiments.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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