Journal: Neuronal Signaling

Article Title: Inhibition of miR-181a promotes midbrain neuronal growth through a Smad1/5-dependent mechanism: implications for Parkinson’s disease

doi: 10.1042/NS20170181

Figure Lengend Snippet: ( A,B ) Neurite length of neurons in E14 rat VM cultures transfected with control siRNA, Smad4 siRNA or Smad1 siRNA or Smad5 siRNA and a GFP-expressing plasmid before being cultured with or without 10 ng/ml GDF5 for 72 h (*** P <0.001 compared with control; ANOVA with post-hoc Tukey’s test; 40 cells for each group per experiment; number of repetitions ( n )=3 experiments). All data are presented as mean ± S.E.M. ( C ) Representative photomicrographs of GFP+ neurons in cultures of E14 rat VM. Scale bar as indicated.

Article Snippet: Where indicated, cells were treated daily with 10 or 200 ng/ml of growth differentiation factor (GDF) 5 (GDF5) (Biopharm GmbH), which are maximal saturating concentrations in primary and SH-SY5Y cell cultures respectively [ , ].

Techniques: Transfection, Control, Expressing, Plasmid Preparation, Cell Culture

Journal: Neuronal Signaling

Article Title: Inhibition of miR-181a promotes midbrain neuronal growth through a Smad1/5-dependent mechanism: implications for Parkinson’s disease

doi: 10.1042/NS20170181

Figure Lengend Snippet: ( A ) Densitometric analysis of p-Smad1/5 in SH-SY5Y cells treated with 200 ng/ml of GDF5 for 0 (control), 5, 30 and 60 min. ( B ) Neurite length of GDF5-treated (200 ng/ml daily for 72 h) SH-SY5Y cells at 72 h. ( C–F ) p-Smad1/5 levels as determined by ( C, D ) Western blots or ( E, F ) immunocytochemistry in SH-SY5Y cells 24 h after transfection with control miRNA or miR-181a inhibitor. Total Smad1/5 was used as a loading control. ( G ) Neurite length, ( H ) neurite branching and ( I ) representative photomicrographs of miR-181a inhibitor or control miRNA transfected SH-SY5Y cells at 72 h (* P <0.05, ** P <0.01, *** P <0.001 compared with control; t test; 40 cells for each group per experiment; n =3 experiments). All data are presented as mean ± S.E.M. Scale bar as indicated.

Article Snippet: Where indicated, cells were treated daily with 10 or 200 ng/ml of growth differentiation factor (GDF) 5 (GDF5) (Biopharm GmbH), which are maximal saturating concentrations in primary and SH-SY5Y cell cultures respectively [ , ].

Techniques: Control, Western Blot, Immunocytochemistry, Transfection