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. 2016 Feb 29;5(2):e201. doi: 10.1038/oncsis.2016.14

Figure 1.

Figure 1

P17 binding and apoptosis analysis in Jurkat cell line after apoptosis induction. (a) Fluorescent microscopy observation of extremely large uptake of FITC-P17 (green) by TRAIL-treated Jurkat cell line. Nuclei stained blue with DAPI. Scale bars, 10 μm (top); 50 μm (bottom). (b) Apoptosis analysis of the same sample used for P17 binding by double staining with annexin V and PI. The quadrants Q were defined as Q1=live (Annexin V- and PI-negative), Q2=early stage of apoptosis (Annexin V-positive/PI-negative), Q3= late stage of apoptosis (Annexin V- and PI-positive) and Q4=necrosis (Annexin V-negative/PI-positive). (c) Flow cytometry quantification of the cellular fluorescence in Jurkat cells treated with TRAIL following incubated with FITC-P17 versus control samples shown via mean fluorescence intensity. Percentage of apoptosis was determined using Annexin V/PI staining and expressed as the proportion of annexin V-positive cell counts in total cell populations. Data are presented as means±s.d. (n=3). ***P<0.001. (d, e) FITC-P17 (green), DAPI (blue) and Annexin V (red; d) or cleaved caspase-3 antibody (red; e) were colocalized within the same Jurkat cell after treatment with TRAIL to induce apoptosis by confocal microscopy. Scale bars, 10 μm (inserts in d); 30 μm (bottom).