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. 2014 Jul 31;5(16):7183–7197. doi: 10.18632/oncotarget.2293

Fig.4. Effect of Apollon knockdown on the chemosensitivity of ESCC cells to cisplatin (CDDP) and docetaxel in vitro.

Fig.4

(A) KYSE510 cells transfected with control shRNA (shC) or two different shRNAs against human Apollon (sh1, sh2) were treated with CDDP (10 μM) for 24 hours, then apoptosis was detected by Annexin V–PI staining. (B) KYSE510 cells and Eca109 cells transfected with control shRNA (shC) or Apollon shRNAs (sh1, sh2) were treated with CDDP (10 μM) or docetaxel (10 nM) for the indicated times. Apoptosis was quantified by annexin V-positive cells. (C) KYSE510 cells transfected with control shRNA (shC) or Apollon shRNAs (sh1, sh2) were treated with CDDP (10 μM) for 24 hours, then Mitochondrial inner transmembrane potential (Δψm) depolarization was detected by Mito Tracker Red staining. (D, E) KYSE510 cells and Eca109 cells transfected with control shRNA (shC) or Apollon shRNAs (sh1, sh2) were treated with CDDP (10 μM) or docetaxel (10 nM) for the indicated times, Δψm depolarization was quantified as the percentage of depolarized cells. Caspase-9 and caspase-8 activation were quantified using a luminescent reporter. All data are represented as Means ± standard deviations (SD) from three to four independent experiments. *P < 0.05 and **P < 0.01 vs. shC. (F) KYSE510 cells and Eca109 cells transfected with control shRNA (shC) or Apollon shRNAs (sh1, sh2) were treated with vehicle, CDDP (10 μM) or docetaxel (10 nM) for 6 hours, long-term cell viability was assessed by the colony formation assay. Representative pictures are shown.