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. 2014 Oct 31;9(10):e111611. doi: 10.1371/journal.pone.0111611

Figure 1. CDKN3 negatively regulates K562 cell survival.

Figure 1

(A) Expression of CDKN3 in K562 cells stably overexpressing FLAG-tagged wild type CDKN3 (CDKN3-WT) or empty vector (EV) was detected by Western blotting using indicated antibodies. (B) K562 cells stably overexpressing CDKN3-WT or EV were treated with 10 µM of imatinib for the indicated time. Samples were stained with Annexin V-APC and PI, examined by flow cytometry and analyzed by FCS Express V3. Plotted are results from three independent experiments. Error bars represent SEM, n = 3; *P<0.05. (C) Shown is an immunoblot examining FLAG-tagged CDKN3-C140S in K562 cells ectopically expressing CDKN3 mutant (CDKN3-C140S) or empty vector (EV). (D) Cell viability of K562 cells expressing CDKN3-C140S or EV was assessed by flow cytometry after treatment with 10 µM of imatinib for 36 h. Samples were stained with Annexin V-APC and PI. Plotted are results from three independent experiments. Error bars represent SEM, n = 3. (E) Shown is an immunoblot examining shRNA-based knockdown of CDKN3. (F) K562 cells stably expressing sh-luc or sh-CDKN3 were treated with 10 µM imatinib for the indicated time. Samples were then stained with Annexin V-APC and PI, followed by flow cytometry analysis. Plotted are results from three independent experiments. Error bars represent SEM, n = 3; *P<0.05.