Skip to main content
. 2016 May 3;107(6):746–754. doi: 10.1111/cas.12937

Figure 4.

Figure 4

Mechanism dissection studies on interleukin‐6 (IL‐6) regulation of hypoxia‐inducible factor (HIF) following cisplatin treatment. (a) Hypoxia response element–luciferase (HRE‐luc) assay. HEK293 cells were transfected with HRE‐luc‐containing plasmids and incubated with various amounts of IL‐6. After 24 h of incubation, luciferase activities were measured. (b) HRE‐luc assay. A549IL‐6si/sc and H157IL‐6si/sc pairs were transfected with equal amounts of HRE‐luc‐containing plasmids. After 24 h of transfection, luciferase activities were measured. (c) Prolyl hydroxylase (PHD) mRNA levels in A549IL‐6si/sc and H157IL‐6si/sc cell sets, with or without cisplatin treatment. Cells were either non‐treated or treated with cisplatin (5 μM, 72 h), total RNAs extracted, cDNA converted, and mRNA expressions were analyzed. (d) Western blot analysis of von Hippel‐Lindau disease tumor suppressor (VHL), HIF1α, and HIF2α levels in A549IL‐6si/sc and H157IL‐6si/sc cell sets, with or without cisplatin treatment, in the presence or absence of MG132. Cells were either non‐treated or treated with cisplatin (5 μM, 72 h) in the absence or presence of MG132 (10 μM), cell extracts were obtained, and HIF1α and HIF2α levels were analyzed in Western blot. (e) Summary of the results of mechanism studies. *P < 0.05; **P < 0.01; ***P < 0.001. CSC, cancer stem cell.