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. 2016 May 26;6:26804. doi: 10.1038/srep26804

Figure 1. KD of Cops2, but not any other CSN subunits, reduces the expression of Nanog and Oct4.

Figure 1

(a) The KD efficiencies of shRNAs targeting CSN subunits. Mouse ESCs were transfected with shRNA plasmids targeting GFP or CSN subunits. Forty-eight hours later, cells were harvested for RNA prep and quantitative RT-PCR. (b) The effect on pluripotency gene expression after KD of CSN subunits. The samples were prepared as described in (a). Quantitative RT-PCR was performed to measure the RNA levels of Nanog, Oct4 and Sox2. (c,d) Two shRNAs targeting Cops2 validate the regulatory effect on Nanog and Oct4. Mouse ESCs were transfected with plasmids expressing shRNAs targeting different sequences of Cops2 (C2-1 and C2-2), GFP or Cops8 (C8). Seventy-two hours later, cells were harvested for quantitative RT-PCR (c) and Western blot (d). The averages (Ave) and standard deviations (SD) of quantified Nanog expression levels from three independent experiments are shown below the blot. (e) Cops2 KD activates the expression of differentiation genes, except for Pax6. Quantitative RT-PCR was performed with the same cells described in (c). Data are shown as mean ± SD (n = 3).