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. 2016 Nov 26;22(2):239–253. doi: 10.1007/s10495-016-1320-8

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© The Author(s) 2016

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

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Fig. 3 — Subcellular distribution of Bax in thymocytes upon DX treatment. a Western blot shows the DX treatment-induced redistribution of Bax between the cytoplasmic and mitochondrial fractions of thymocytes. Blots were reprobed with anti-β-actin or anti-Cytochrome C (Cyt C) antibodies to confirm the purity of the cytoplasmic and mitochondrial fractions, respectively. The figure shows a representative blot and the densitometry data of at least three independent experiments. The diagram shows the relative Bax expression in the cytoplasm (normalized to β-actin) and the mitochondria (normalized to Cytochrome C). Bars represent the mean ± SEM of relative densities compared to the controls. b Mitochondrial translocation of Bax in DP thymocytes. b1 Representative confocal microscopic images of at least three independent experiment showing CMX-Ros-Bax co-localization in control (Ctrl) and 30 min DX-treated cells. DIC, CD4 (blue channel) and CD8 (virtual red channel) overlaid, mitochondria (CMX-Ros, red channel) and Bax (green channel) images are shown. The co-localization of mitochondria with Bax (CMX-Ros-Bax merged images) is indicated by yellow areas. Scale bars are 8 µm each. b2 Quantification of the changes in the CMX-Ros-Bax co-localization in DP thymocytes after in vitro DX treatment was performed using the co-localization plugin of the ImageJ software. Bars represent the number of co-localized pixels. The mean ± SEM was calculated from the data of 100 DP cells per treatment, respectively. (Color figure online)