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Fig. 3 | Journal of Hematology & Oncology

Fig. 3

From: Adenylate kinase 4 modulates oxidative stress and stabilizes HIF-1α to drive lung adenocarcinoma metastasis

Fig. 3

AK4 stabilizes and exaggerates HIF-1α protein expression to promote EMT. a Upper, endogenous AK4 and HIF-1α protein expression in human NSCLC cell lines. Bottom, correlation between AK4 and HIF-1α protein expression in NSCLC cell lines. b Left, WB analysis of AK4, HIF-1α, E-cadherin, and vimentin upon AK4 knockdown in CL1-5 cells. Right, WB analysis of AK4, AMPK, phospho-AMPK (Thr172), and E-cadherin upon AK4 knockdown in A549 cells treated with or without TGF-β (5 ng/mL) for 24 h. c WB analysis of HIF-1α, AK4, E-cadherin, vimentin, and Snail in CL1-0 and H1355 vector- or AK4-overexpressing cells exposed to hypoxia for the indicated time. d RT-PCR analysis of AK4, HIF1A, CDH1, HK2, GLUT1, VEGFA, and GAPDH in CL1-0 vector- or AK4-expressing cells under normoxic and hypoxic conditions. e WB analysis of HIF-1α from CL1-0 vector- or AK4-expressing cells treated with the proteasome inhibitor MG-132 under Nx and Hx conditions. f WB analysis of HIF-1α and AK4 from CL1-0 vector- or AK4-expressing cells treated with CHX for 20, 40, and 60 min. g WB analysis of HIF-1α and hydroxylated HIF-1α from CL1-0 vector or AK4-expressing cells treated with MG-132 under Nx. h Invasion assay of CL1-0 vector- or AK4-expressing cells under normoxia (Nx) or hypoxia (Hx). **P ≤ 0.01. i Invasion assay of H1355 vector- or AK4-expressing cells under Nx or Hx. **P ≤ 0.01. The results are presented as the mean ± SD of at least three separate experiments. Two-tailed, unpaired Student’s t tests were used for all pairwise comparisons. **P ≤ 0.01

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