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. 2018 Sep 13;8:13790. doi: 10.1038/s41598-018-31696-9

Figure 5.

Figure 5

GRG5 is essential for the direct conversion of MEFs to induced neurons. (a) Schematic representation of the experimental process for the direct neuronal conversion of MEFs. MEFs were infected with inducible lentiviruses expressing the proneural factors ASCL1 and MYT1L. Silencing of GRG5 expression was achieved using specific shRNA. From Day 2 post-infection cells were cultured in neuronal promoting conditions (iN medium). On Day14 the generation of induced neuronal cells (iNs) was evaluated. (b) Representative immunofluorescence photos showing TUJ1 positive neuronal cells derived from CONTROL and KD GRG5 MEFs at Day 14. (Scale bar, 50 μm) (c) Conversion efficiency estimated by dividing the number of TUJ1 positive cells at Day14 with the number of the plated MEFs at Day 0. Mean + SD of n = 3 independent experiments. *P < 0.05 (d) Neuronal purity defined as the number of TUJ1 positive cells at Day14 divided by the total number of DAPI positive cells at Day 14. Mean + SD of n = 3 independent experiments. *P < 0.05 (e) Luciferase assay monitoring the activity of BMP signaling using the BRE-LUC reporter plasmid in MEFs upon transient silencing or over-expression of GRG5. Mean + SD of n = 3 independent experiments. *P < 0.05.