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. 2015 Feb 17;12(1):83–92. doi: 10.3892/mmr.2015.3368

Figure 4.

Figure 4

SOCS3 inhibits HIF-1α expression through the Akt pathway but not the STAT3 pathway. Protein levels of HIF-1α, STAT3, Akt and the phosphorylated, activated forms pSTAT3 and pAkt were assessed using western blot analysis. Representative images of three independent experiments and quantified levels normalized to β-actin are shown. (A) Following transfection of NCI-H446 cells with Ad5-SOCS3, the expression levels of HIF-1α, STAT3, Akt as well as pSTAT3 and pAkt decreased significantly (P<0.05, Ad5-SOCS3 group vs, control group). (B) In NCI-H446 co-transfected with Ad5-SOCS3 and pcDNA3-STAT3, STAT3 and pSTAT3 expression were upregulated (*P<0.05), while HIF-1α expression levels exhibited no significant change (▲P>0.05). However, when co-transfected with pcDNA3-Akt not only Akt and pAkt, but also HIF-1α expression levels were upregulated (*P<0.05). (C) In NCI-H446 cells treated with the STAT3 inhibitor CPA-7, HIF-1α expression levels exhibited no significant change (▲P>0.05). By contrast, when treated with Akt inhibitor wortmannin, HIF-1α expression levels were markedly inhibited (*P<0.05). SOCS3, suppressor of cytokine signaling 3; HIF-1α, hypoxia-inducible factor-1α; STAT3, signal transducer and transcription activator 3; p, phosphorylated.