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. 2017 Sep 7;10:283. doi: 10.3389/fnmol.2017.00283

Figure 1.

Figure 1

Lentiviral-mediated knockdown of Forebrain embryonic zinc finger 2 (Fezf2) in the mature motor cortex. (A) Male mice aged P21–25 were injected into the primary motor cortex (M1) with lentivirus (LV) expressing a mCherry reporter gene and either a Fezf2 shRNA or non-silencing shRNA. Four weeks post-injection M1 tissue was isolated according to mCherry expression and processed for RNA-seq analysis. (B) Schematic indicating the binding site for the Fezf2 shRNA in the Fezf2 mRNA. (C) Injection of LV.Syn.mCherry.non-silencingshRNA in M1 of Fezf2-Gfp mice validated the successful transduction of Fezf2-expressing neurons. Brain section is from Bregma +0.4. Scale bar is 500 μm or 50 μm (higher magnification images). (D) Quantitative PCR analysis was used to test the shRNA-mediated knockdown of Fezf2 expression in the mature M1 of wildtype male mice (n = 4, 4; *p < 0.05, unpaired one-tailed t-test). (E) Stable HT1080 cells transduced with a green fluorescence protein (GFP)-mFEZF2 fusion construct were induced to express either the Fezf2 shRNA or non-silencing shRNA (control). Images indicate shRNA expression according the RFP reporter and GFP-mFEZF2 expression (GFP) at 0 h and 72 h after the induction of shRNA expression. Scale bar is 100 μm.