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Fig. 4.

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Fig. 4. Depletion of HYOU1 inhibits tumor growth in lung cancer cells. (A) Expression levels of HIF1 and HYOU1 in monolayer (2D)- or spheroid (3D)-cultured NSCLC cells (NCI-H460, A549, H1299, and PC9), as assessed by western blot analysis. (B) Clonogenic survival in NSCLC cells (NCI-H460, A549, and H1299) transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1), as assessed by colony formation assay. (C) Immunofluorescence and bright-field images of lung cancer spheroids (NCI-H460, A549, and H1299) transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1). The spheroids were stained with 4 μM EthD-1. The images were obtained using the Operetta® High Content Screening System, and the intensity of EthD-1 staining in lung cancer spheroids relative to controls was analyzed using Harmony software. (D) Expression of cleaved caspase-3, cleaved PARP, HIF1, HYOU1, pp38, p53, and pErk1/2 in lung cancer spheroids (NCI-H460 and A549) transfected with nonspecific siRNA (siCont) or HYOU1 siRNA (siHYOU1). The data shown are the mean ± SD from three independent experiments; **P < 0.01, ***P < 0.001, and ****P < 0.0001 compared to the control group.
Mol. Cells 2021;44:50~62 https://doi.org/10.14348/molcells.2020.0212
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