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. 2016 Sep 12;10:217. doi: 10.3389/fncel.2016.00217

Figure 4.

Figure 4

Recombinant hGDNF (rhGDNF) exogenously added at early stages increases differentiation towards MN, which can be prevented by neutralizing anti-GDNF antibodies. CTRL-ESC/HB9::GFP were differentiated without rhGDNF (A) or adding this protein only at specific periods: prior to formation of EBs (B), during EBs (C) or after EBs (D). The quantification of total MN differentiation (ISL1 and/or GFP) reveals that early and transient exposure to rhGDNF significantly increases MN induction (E). **p < 0.001 and ***p < 0.0001 compared to differentiation without rhGDNF. Addition of anti-GDNF neutralizing antibodies did not affect differentiation of CTRL-ESC/HB9::GFP (F). However, co-incubation of rhGDNF and anti-GDNF antibodies, significantly decreased the inductive effect of GDNF. **p < 0.001 and ***p < 0.0001 compared to differentiation with rhGDNF. Plot resulting from three independent experiments performed in duplicate. Scale bar: 50 μm.