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. Author manuscript; available in PMC: 2014 Jul 3.
Published in final edited form as: Cell. 2013 Jul 3;154(1):118–133. doi: 10.1016/j.cell.2013.05.059

Figure 4. α1ACT enhances neurite outgrowth by regulating BTG1 expression.

Figure 4

(A) Relative mRNA expression levels of BTG1, PMCA2, TAF and GRN in PC12 cells transfected with α1AWT, α1ACTWT or α1ACTSCA6 (n ≥ 3). (B and C) Western blot (B) and quantitation of protein expression levels of BTG1 and PMCA2 in PC12 cells transfected with α1AWT, α1ACTWT and α1ACTSCA6 (C). (D) Relative levels of BTG1 mRNA in the cerebellum from two SCA6 patients, normalized to Pcp2. (E and F) α1ACTWT enhances neurite outgrowth. Representative low- and high-magnification images of PC12 cells with transiently transfected pcDNA3-FLAG, α1AWT-FLAG, α1ACTWT-FLAG and α1ACTSCA6-FLAG at 24 hr (E) and 72 hr after NGF treatment (F). Cells were labeled for GAP-43 (green) to visualize PC12 cell body and neurites. (G and H) Quantitation of average neurite length and percentage of neuritis per cell (n = 200; *p<0.05 versus pcDNA3-FLAG). (I) α1ACTWT up-regulates BTG1 gene and increases PRMT1/BTG1 protein interaction. (J and K) Silencing of BTG1 expression inhibits α1ACTWT-enhanced neurite outgrowth. Anti-FLAG staining is shown in red. (L) Quantitation of neurite outgrowth by siBTG1 in transfected cells (n = 3, *p<0.05). The blunted effect by α1ACTSCA6-FLAG was also diminished by BTG1 silencing. Data are mean ± SEM (see also Figure S4).