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. 2020 Sep 1;31(19):2125–2138. doi: 10.1091/mbc.E20-04-0237-T

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© 2020 Hertzler et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology.

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FIGURE 5: — DLK-mediated cell stress is not responsible for the microtubule phenotype caused by kinetochore knockdowns. (A) Kymographs of EB1 videos taken of ddaE neurons in larvae treated with GNE-3511, a DLK inhibitor. Compared to DMSO vehicle controls, GNE-3511 treatment did not result in a significant suppression of the microtubule dynamics phenotype (B) induced by Ndc80, Nuf2, or Bub3 RNAis. Statistics are nonparametric, Mann–Whitney t tests between DMSO and GNE-3511 treatments for each genotype separately. Numbers in columns are number of neurons analyzed. (C) puc, a negative regulator of the DLK cell stress pathway, is expressed as a function of DLK activation and accumulates in the nucleus, visible with the puc-GFP construct. Pictured are example images of control (left), severe DLK activation by unc-104 RNAi (middle), and Ndc80 RNAi (right). mCD8-RFP is used as a cell shape marker. (D) The average fluorescence intensity of puc-GFP in the nucleus is measured for each cell and normalized to the control average, which is set to 1. **, p < 0.01; *, p < 0.05, with a Kruskal–Wallis one-way ANOVA, each genotype compared with the control with Dunn’s multiple comparisons test. Error bars are SD.