MEF cells lacking TIA proteins show mitochondrial alterations.

<p>(A) Wild-type (WT), TIA1 KO and TIAR KO MEFs were grown for 4 days and monitored by methyl thiazolyl tetrazolium (MTT) assays. The represented values were normalized and expressed relative to WT, whose value was fixed arbitrarily to 1, and are means + SEM (n = 4; *<i>P</i><0.001). (B) Wild-type (WT), TIA1 KO and TIAR KO MEFs were stained with Mito Tracker Green FM and monitored by FACS analysis. The represented values are means + SEM (n = 3; *<i>P</i><0.01; **<i>P</i><0.001). (C) Mitochondrial populations of wild-type (WT), TIA1 KO and TIAR KO MEF were stained with Mito Tracker Green FM and visualized by confocal microscopy. Scale bars represent 10 µm. (D) Mitochondrial DNA copy number was estimated by quantitative PCR (QPCR) using mitochondrial NADH subunit 1 (ND1) and cytochrome c oxidase subunit 1 (CO1) and nuclear tumor necrosis factor (TNF) and H19 as markers for the copy numbers of mitochondrial DNA (mtDNA) and nuclear DNA (nDNA), respectively. The represented values were normalized and expressed relative to WT, whose value was fixed arbitrarily to 1, and are means + SEM (n = 4; *<i>P</i><0.05). (E) Estimation of the mitochondrial membrane potential of wild-type (WT), TIA1 KO and TIAR KO MEFs by staining with TMRM molecular probe and monitoring by FACS analysis. The represented values are means + SEM (n = 3; *<i>P</i><0.001). (F) Estimation of reactive oxygen species (ROS) of wild-type (WT), TIA1 KO and TIAR KO MEFs by staining with H2DCFDA molecular probe and monitoring by FACS analysis. The represented values are means + SEM (n = 3; *<i>P</i><0.01). (G) Transmission electron micrographs of cytoplasmic sections from wild-type (WT), TIA1 KO and TIAR KO MEFs illustrating the mitochondrial morphology and integrity. Scale bars represent 400 nm.