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. 2016 May 5;7(24):36814–36828. doi: 10.18632/oncotarget.9192

Figure 2. AR is co-localization with Nanog in HCC cells based on GFP labeled Nanog cells by CRISPR/Cas9 system.

Figure 2

A. The schematic of the Nanog labeled CRISPR/Cas9 system: the green capital refers to Cas9 recognize protospacer adjacent motifs (PAM) sequence, and the red lowercase refers to Nanog termination codon. Nanog exon4 without termination code jointed with 2A-GFP sequence, and each length of homologous arm is 2Kb. B-C. RT-qPCR and Western blot were performed to measure the mRNA of Nanog in GFP (+)/(−) cells of two labeled clone cells, qPCR values are normalized to GAPDH and represent the mean ± SD of triplicate samples. D. The confocal image of Nanog and GFP co-localization in T1224+1 and Huh7+7 cells, blue: DAPI, green: GFP, red: NANOG, white arrow: co-localization positive, red arrow: co-localization negative. E-F. RT-qPCR and Western blot detected the expression of Nanog and AR in two clone GFP (+)/(−)cells. G. The confocal images of T1224+1 and Huh7+7, blue: DAPI, green: GFP, red: AR, white arrow: co-localization positive, red arrow: co-localization negative. Scale bars, 100μm, p<0.05(*), p<0.01(**) and p<0.001(***).