Supplementary Figure 1: PTB knockdown in human adult fibroblasts dampens cell proliferation and cell fate switch to the neuronal lineage.
From: Sequential regulatory loops as key gatekeepers for neuronal reprogramming in human cells
(a) Characterization of HAFs by immunostaining with various cell type markers. HAFs show homogeneous staining of the fibroblast markers Fibronectin, FSP1 and Vimentin, but not markers for keratinocytes (K5, P63), melanocytes (MITF, P75), neural crest derivatives (SOX10, BRN3A) and neurons (indicated in the lower panel). n=2 biological repeats.
(b) FAC analysis of EdU-pulse labeled HAFs with or without PTB knockdown. Nuclei were stained by DAPI and EdU positive cells were stained with Alexa fluor 647. n=3 biological repeats.
(c) Example of Tuj1 (Green) positive cells that lack EdU (Red) staining. n=3 biological repeats.
(d) Example of induced neuronal-like cells by PTB knockdown only or in combination with specific small molecules. Red: Tuj1 expression, Blue: DAPI-stained nuclei. Scale bar: 20 µm. n=2 biological repeats.
(e) Phase contrast pictures of control and PTB shRNA-treated HAFs with small molecules treatment at different time points. The 3 small molecules were either present through the assay periods or washed away from the first treatment for 3 hrs. n=2 biological repeats.
(f) Immunostaining of the neural crest markers Nestin and Sox2 during PTB knockdown induced neuronal conversion. Scale bar: 100 µm. n=3 biological repeats.
