Mitochondrial network in Apaf1-deficient cells is affected.

<p>(A) Bcl-2 and Bcl-X_L protein levels in the SV40IM and SIM cell lines by immunoblotting. α-tubulin was also assayed as a loading control. Densitometric analyses were performed using the Image J software, normalised for α-tubulin and reported as arbitrary units (a.u). Values are means ± s.d. of three independent immunoblots. (B) The mitochondrial network in the untreated SV40IM and SIM MEF cell lines was visualised using an anti-Cyt <i>c</i> antibody. (C) Transmission electron photomicrographs of the mitochondria in MEFs. Ultrathin (0.07 µm) sections of the MEF cell lines were prepared as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084666#s2" target="_blank">sMaterials and Method</a>. Black arrows indicate the representative mitochondria of each sample. Scale bars indicate 1 µm. The Image J Java-based image processing software was used to quantify the width of approximately 70 different mitochondria from each cell type using the images obtained by TEM. Two-tailed Student's t-tests were performed (***<i>p</i>≤0.001). SV40IM-MEFS WT Apaf1 is of 0.2939±0.01501 µm and SV40IM-MEFS KO Apaf1 is of 0.1980±0.009215 µm. SIM-MEFS WT Apaf1 is of 0.3539±0.01625 µm and SIM-MEFS KO Apaf1 is of 0.1697±0.02228 µm.