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. 2016 Jun 9;7(6):e2260. doi: 10.1038/cddis.2016.162

Figure 7.

Figure 7

Thymocyte analysis in Mysm1−/− mice. (a) Thymocytes were isolated from 8- to 10-week-old WT or Mysm1−/− mice (n=4) for preparation of cell lysates. The cell lysates were fractionated by 4–12% SDS-PAGE and analyzed by WB to determine IRF2 expression. (b) Thymocytes were isolated from 8- to 10-week-old WT or Mysm1−/− mice (n=4) and analyzed via immunostaining/FACS using anti-CD4, anti-CD8, and anti-Sca1 antibodies to determine Sca1 expression in the different stages of thymocyte development. (c) Thymocytes from 8- to 10-week-old WT or Mysm1−/− mice (n=4) were used to prepare cell lysates. The cell lysates were analyzed via WB for detecting expression of pro-apoptotic proteins Bax/Bak and anti-apoptotic proteins Bcl-2/Bcl-x. (d) Thymocytes from 8- to 10-week-old WT or Mysm1−/− mice (n=4) were analyzed for ROS levels (left) and for mitochondrial membrane potential (right). (e) The isolated thymoctyes were cultured overnight under the indicated conditions and analyzed via Annexin V/PI staining to define cell death. Data are representative of three independent experiments. DN, double-negative thymocyte; DP, double-positve thymocyte; SP, single-positive thymocyte