Lmo4 synergizes with Fezf2 to promote direct in vivo reprogramming of upper layer cortical neurons and cortical glia towards deep-layer neuron identities
Fig 2
Reprogramming of ULs into layer V neurons by Fezf2 and Lmo4 is more efficient in motor than somatosensory cortex.
(A) Schematic representation of the experimental procedure. cGFP, cLmo4 (cL), cFezf2 (cF) or cFezf2 and cLmo4 (cF+cL) plasmids were electroporated into E14.5 motor (M1) cortices. Brains were collected at P7. (B) IF of GFP, UL marker Cux1, and DL V marker Ctip2 on a coronal slice of a cF+cL-electroporated brain. The white box indicates the magnification image on the right side. (C) Percentage of M1 electroporated-UL neurons expressing UL vs. DL markers. (D, E) Percentage of S1 vs. M1-electroporated UL neurons expressing layer V marker Ctip2 (D) and layer VI marker Fog2 (E). Scale bars: B = 1,000 μm (left, macro image), 200 μm (right, magnified image). Results are represented as mean ± SEM. Two-way ANOVA with Tukey’s post hoc correction (2C) or two-way ANOVA with Sidak’s post hoc correction (2D-E) was used for statistical analysis. *p < 0.5, **p < 0.01, ***p < 0.001, ns = not significant. n = 3 brains for each plasmid. Extended data and statistics are listed in S2 Data. DL, deep layers; GFP, green fluorescent protein; IF, immunofluorescence; UL, upper layers.