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GSK-3α knockdown suppresses lung cancer cell viability.

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posted on 2016-04-06, 04:51 authored by Sin-Aye Park, Jong Woo Lee, Roy S. Herbst, Ja Seok Koo

(A) Effect of CREB knockdown on the cell viability. Lung cancer cell lines (H1993, H1437, H1734, and A549) were transiently transfected with control siRNA (50 nM), or CREB siRNA (10, 50 nM) for 72 h, followed by MTT assay. All values in the graphs represent mean ± SD of three independent experiments. Two-sided t-test. *, P < 0.01. (B) Effect of GSK-3α knockdown on the cell viability. The cells were transiently transfected with control siRNA, or GSK-3α siRNA (40 nM, each) for 72 h and the quantitative data was followed by MTT assay. All values in the graphs represent mean ± SD of three independent experiments. Two-sided t-test. *, P < 0.01. (C) Effect of GSK-3α knockdown on colony formation. One day after transfection of control siRNA (40 nM), or GSK-3α siRNA (10, 40 nM), the cells were seeded again in 6-wells with low density (2 x 103/ well) and incubated for 7–14 days. Mean ± SD in three independent experiments. Two-sided t-test. *, P < 0.01. (D) Effect of GSK-3α knockdown on the cell death. H1734 and H1993 cells were infected with lentiviral expressing shRNA targeting GSK-3α (two sequences; shGSK3A#2 and shGSK3A#4) with 8 μg/ml polybrene. After 48 h infection, cells were selected with 0.5 μg/ml puromycin for 3 days. The selected cells were performed by staining of annexin V and PI to analysis apoptotic cell death by LSRII.

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