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. Author manuscript; available in PMC: 2014 Jul 16.
Published in final edited form as: Cancer Res. 2013 Apr 11;73(8):2650–2658. doi: 10.1158/0008-5472.CAN-12-3158

Figure 3.

Figure 3

Loss of PRMT5 results in elevated Gas1 levels and enhanced Hedgehog (Hh) signaling. (A) Quantitative RT-PCR (qRT-PCR) showing reduction of Prmt5 mRNA in MEFs expressing two independent Prmt5-targeting shRNA clones compared to MEFs expressing scrambled control shRNA. (B) qRT-PCR showing expression of Gas1 mRNA levels in MEFs expressing shRNA targeting Prmt5. (C) Immunoblotting for PRMT5 and GAS1 in MEFs expressing either control or Prmt5-targeting shRNA’s. Ponceau S is included as a loading control. (D) Chromatin immunoprecipitation (ChIP) with antibodies against PRMT5 and histone H4 arginine 3 symmetric dimethylation (H4R3m2s) at the Gas1 promoter. ChIP amplicon, Gas1 (−780 bp / −609 bp). (E–F) MEFs expressing control or two independent shRNA clones targeting Prmt5 were cultured in either control or Sonic hedgehog conditioned medium (Shh-CM) and Gli1 (E) and Ptch1 (F) mRNA levels were quantitated by qRT-PCR. Error bars indicate ± s.d.