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. 2015 Jun 8;6(27):23496–23509. doi: 10.18632/oncotarget.4360

Figure 2. HOXC8 is involved in embigin transcription in MDA-MB-231 and MCF7 cells.

Figure 2

A. MDA-MB-231 and MCF7 cells were lentivirally transduced with empty vector or vector encoding HOXC8, and then transfected with EMB promoter luciferase reporter vectors that were generated using PCR amplification. Luciferase activity was measured 24h posttransfection and normalized using Renilla activities. Columns, mean; bars, SEM; *, P < 0.05. B. MDA-MB-231 and MCF7 cells were lentivirally transduced with scrambled shRNA or HOXC8 shRNA, and then transfected with EMB promoter luciferase reporter vectors along with Renilla for normalization. Luciferase activity was measured 24h posttransfection. Columns, mean; bars, SEM; *, P < 0.05. C. MDA-MB-231 or MCF7 cells were lentivirally transduced with empty vector or HOXC8 expression vector, and then were treated with 2μg/ml actinomycin for different time. Total RNA was isolated and subjected to qRT-PCR to measure the level of EMB mRNA. β-actin and GAPDH mRNA were used as internal controls. The level of EMB mRNA without actinomycin treatment was considered as 100%. Values are means ± SEM; n = 3. D. MDA-MB-231 or MCF7 cells transfected with scrambled or HOXC8 shRNA were treated 2μg/ml actinomycin. Total RNA was isolated at varying times and then subjected to qRT-PCR to measure the level of EMB mRNA. GAPDH and β-actin mRNA were used as internal controls. The level of EMB mRNA without actinomycin treatment was considered as 100%. Values are means ± SEM; n = 3.