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. 2012 Apr 4;103(6):1028–1037. doi: 10.1111/j.1349-7006.2012.02260.x

Figure 5.

Figure 5

Inhibitor of DNA binding protein 2 or 4 (Id2 and Id4) overexpression induces the downregulation of Olig1 and Olig2 transcription factors in glioblastoma multiforme (GBM) neurospheres. (A) HSR‐GBM1B and HSR‐GBM1A cells were transfected with Id2 or Id4 cDNA for 72 h. Id2 and Id4 overexpression downregulated Olig1 and Olig2. (B) Immunofluorescent stainings confirmed that intracellular levels of Olig1 and Olig2 were decreased in Id2 or Id4 overexpressing GBM neurosphere cells. Z‐stack fluorescent images were taken using fluorescent microscopy. Stains from one single plane of the cells are shown. Olig1 was expressed in both cytoplasm and nuclei; arrows indicate stained nuclei. Olig2 was mainly found in nuclei. Bar = 15 μm. (C) Expression of Olig1 and Olig2 mRNA after Id protein overexpression as detected by quantitative RT‐PCR. Id proteins induced a significant decrease in Olig1 and Olig2 expression at the mRNA level. *P < 0.05; **P < 0.01. (D) Knockdown of Olig1 or Olig2 in GBM neurosphere cells with siRNA. Olig1 or Olig2 expression was decreased by approximately 50–70% after siRNA (SiOL1 or SiOL2) were transfected in GBM neurosphere cells for 72 h.