Fig. 6

OLIG2 is involved in regulation of its own expression at the transcriptional level and after transcription. a Experimental overview. b ChIP-Seq result shows that OLIG2 binding to its promoter region, potentially self-regulates its own transcription. c Binding of OLIG2 to Olig2 mRNA in NSCs shown by RIP-Seq. IgG was used as control. d In vitro transcribed biotinylated Olig2 RNA retrieves OLIG2 protein. The profiles were established by RNA protein pull-down using NSCs protein extract. Retrieved OLIG2 protein (32 kDa) was detected by Western blot assay. e Addition of the Olig2 3′ UTR to the ZsGreen1 reporter C1 end substantially reduces expression of ZsGreen1 in Olig2 overexpressing cells. Flow cytometric analyses on MFI of ZsGreen1 fluorescence signal in 293FT cells 48 h after transfection. The MFI of ZsGreen1 fluorescence signal was normalized by dividing the MFI of mRuby fluorescence signal in each replicate. The error bars show standard errors of the means (error bars) (n = 6 in each assay) f. The normalized protein level of OLIG2 in NSC cells 48 h after transfected ZsGreen1 reporter constructs with or without Olig2 3′ UTR. The protein level of OLIG2 was examined by Western blot and normalized to corresponding GAPDH levels. All quantified data were presented as mean ± SEM (standard error of the mean). N = 3; t test analysis *, p < 0.05; **, p < 0.01