Figure 2. (See previous page).

Direct conversion of MEFs into neuronal cells by BAM in the presence of cell division inhibitors. (A) BrdU incorporation in MEFs treated with cell division inhibitors aphidicolin or mimosine for 24 hr. (B) Number of MEFs survived after 2–10 d of culture in the presence of inhibitors. Aph – aphidicolin, Mim – mimosine, Ctr - untreated cells. X-axis indicates experimental time-frame starting from plating of MEFs (day -2) until day 10. Y-axis indicates the number of survived cells. The arrow shows the moment when medium was supplemented with cytostatics (day 0). The culture medium used at each time point of the experiment is shown in boxes under the plot. (C) Experimental design. MEFs were plated at day -2, transduced with BAM+RT viruses at day -1 and cultured in the presence of DOX and aphidicolin starting from day 0 until day 11. (D) Immunostaining with Tuj1 (yellow) and MAP2 (red) antibodies in the aphidicolin treated cells 5–11 d after viral transduction. (E) Tuj1/NF200 double-stained neurons generated from MEFs in the presence of aphidicolin 10 d after viral transduction. (F) Quantification of synapsin (Syn1) and Tuj1 expression (Log₁₀ scale) in the aphidicolin treated cells 5–11 d after viral transduction. MEFs: control fibroblasts not transduced with viruses and cultured for 11 d in N2B27 media without aphidicolin *P < 0.05 in comparison to control MEFs.