Fig 2: The effectiveness of modified reprogramming protocols. (A) Images illustrating morphology of induced neuron at 0 DIV, 7 DIV, 14 DIV, and 35 DIV. Light microscopy. Scale bars correspond to 100 µm. (B) Representative images of fibroblasts stained with DAPI without transduction of lentiviruses (w/o), after 1 selection with antibiotics, and after re-plating on coverslips and secondary selection with antibiotics, according to the modified protocol. Scale bar corresponds to 50 µm. (C) Representative images of iSNs reprogrammed according to the modified protocol and stained with antibodies to the neuronal markers and counterstained with DAPI. Immunofluorescent staining of induced neurons with antibodies for the neuronal marker protein TUJ1 (secondary antibodies conjugated with the Alexa 488 fluorophore), MAP2 (secondary antibodies conjugated with the Alexa 594 fluorophore), and DARPP-32 (secondary antibodies conjugated with the Alexa 488 fluorophore). Scale bar corresponds to 50 µm. (D) Histogram represents the number of remaining cells without transduction of lentiviruses (w/o) after the first and second steps of selection. Data are presented as the mean ± SEM. n > 500 cells. (E) Histogram represents the number of the TUJ1-, MAP2-, DARPP-32-positive cells counterstained with DAPI and reprogrammed according to the modified protocol. Data are presented as the mean ± SEM. n > 250 cells for each staining.