1. Fig. 4 IDH3α interacts with cSHMT during S phase at the nuclear lamina.
IDH3α interacts with cSHMT during S phase at the nuclear lamina. (A) Cell cycle distribution in unsynchronized and S phase–arrested NHA cells as indicated by propidium iodide staining and flow cytometry–based quantification of G1, G2-M, and S phase content. un, unsynchronized; S, synchronized. (B) WB analysis of IDH3α in heavy membrane (HM) and cytosolic (C) compared to nuclear (Nuc) fractions. Histone H3 and cytochrome c oxidase subunit IV (COXIV) are shown as nuclear and mitochondrial markers, respectively. (C) Confocal IF images of unsynchronized (a and b) and S phase–arrested NHAs (c) stained for IDH3α (green), cytochrome c (cyto c; red), and DNA (Hoechst; blue). Scale bars, 20 μm (a), 14 μm (b), and 22 μm (c). Arrows point to IDH3α colocalizing with cytochrome c, and arrowheads point to IDH3α associated with the nuclear lamina. (D) Venn diagram illustrating IDH3α co-immunoprecipitated proteins using IgG control or IDH3α-specific antibodies in shScr- and shIDH3α-expressing GICs. (E) Bar graph illustrating total normalized cSHMT spectra in IgG and IDH3α immunoprecipitates in the indicated cell lines. (F) IP-WB analysis of IDH3α and cSHMT validates the IDH3α-cSHMT complex.
Jasmine L. May et al. Sci Adv 2019;5:eaat0456
Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).