Article
Extended Data Fig. 4 | Cre-dependent expression of RFP in injected mouse
midbrain. a, Schematic of the substantia nigral region (white box) for A AV
injection and immunochemical analysis. b, Cre-dependent RFP expression.
RFP+ cells were not detected in midbrain of wild-type mice injected with either
A AV-empty or A AV-shPTB (left). In comparison, both viruses generated
abundant RFP signals in Gfap-cre transgenic mice. Scale bar, 150 μm.
c, d, Co-staining of RFP+ cells with the astrocyte markers S100b and ALDH1L1
10 weeks after injecting A AV-empty (c), indicating that most RFP+ cells in
AAV-empty-transduced midbrain were astrocytes. Scale bar, 25 μm. d, No RFP
expression was detectable in NG2-labelled cells. Scale bar, 15 μm. Experiments
in b–d were independently repeated three times with similar results.
e, Reprogramming-dependent conversion from astrocytes to neurons.
Immunostaining with the astrocyte marker GFAP and the pan-neuronal marker
NeuN was performed 10 weeks after injection of A AV-empty or A AV-shPTB in
the midbrain. Scale bar, 30 μm. Quantified results show that cells transduced
with A AV-empty were all GFAP+ astrocytes, whereas cells transduced with
A AV-shPTB were mostly NeuN+ neurons. Quantified data were based on three
mice as shown on the right. Two-sided Student’s t-test. Data are mean ± s.e.m.
f, g, Further characterization of A AV-shPTB-induced neurons in midbrain with
additional neuronal markers, including pan-neuronal specific markers TUJ1,
MAP2, NSE and PSD95 (f; scale bar, 10 μm) and specific markers for
glutamatergic (VGlut2) and GABAergic (GAD65) neurons (g; scale bar, 20 μm).