Fig. 6
From: Binding of HMGN proteins to cell specific enhancers stabilizes cell identity
HMGN depletion enhances direction lineage conversion of MEFs to induced neurons. a Protocol for direct lineage conversion of MEFs to induced neurons by transcription factor ASCL1. b Western blot (left) and qPCR gene expression analysis (right, 3 biological replicates, scale bar: mean ± SD) showing equal expression levels of ASCL1 in WT and DKO MEFs two days after doxycycline induction. For qPCR gene expression analysis, average of three biological replicates is shown and bar graph represents mean ± SD. c Immunofluorescence staining of ASCL1 and TUJI in MEFs at day 0 and day 12 of doxycycline induction. Scale bar: 50μm. d Immunofluorescence staining of ASCL1 and TUJI in WT and DKO cells after 12 days of differentiation induction. Scale bar: 200μm. e Quantitative analysis of neuronal induction efficiency in WT and DKO MEFs by counting the number of TUJ1 positive cells relative to the number of ASCL1 positive cells at different induction time points (average of three biological replicates is shown, mean ± SD). f Western blot analysis of TUJ1 expression in WT and DKO MEFs at different induction time points. g Enhanced expression levels of neuron-specific markers, Tubb3, Nestin and Map2markers in transdifferentiated DKO MEFs. Gene expression level at each time point was normalized to Gapdh expression level (Average of three biological replicates are shown, mean ± SD). T-test was used in e, g. p values: *p < 0.05; **p < 0.01
