Ectopic expression of OSKTFs in retrogradely labeled corticospinal neurons (IMAGE)
Caption
(A) Schematic diagrams of the experimental design and the Tet-on system. (B, C) Representative images of transverse brain sections showing retrogradely labeled corticospinal neurons in mice treated with saline or doxycycline (Dox). Scale bar: 100 μm. (D–K) Representative high-magnitude images of layer V sensorimotor cortex showing fluorescent in situ hybridization of tdTomato and transcriptional factors Oct4, Sox2, Klf4, and Fezf2 in mice treated with saline or Dox. Scale bar: 50 μm. (L–O) Quantification of colocalization between tdTomato and Oct4/Klf (L), Sox2 (M), Fezf2 (N), and tdTomato neuronal densities (O) in animals treated with saline (n = 3) or Dox (n = 3). ∗∗P < 0.01; n.s., no statistical significance; Student's t-test. (P, Q) Neonates (P1) or adult (18 weeks) animals received intraspinal AAVRetro-tdTomato or intersectional injection for OSKTFs expression in corticospinal neurons. Mice were euthanized at P7 or 28 weeks with an 8-week Dox or saline treatment (P). Layer V of the sensorimotor cortex was microdissected under an epi-fluorescent microscope (Q) for measuring RNA contents. Scale bar: 500 μm. (R) Relative RNA expression of different histone proteins and epigenetic modulators in P7 mice (n = 5 biological replicates) and adult mice treated with saline (n = 5 biological replicates) or Dox (n = 5 biological replicates). RNA expression levels were first normalized to those of GAPDH and then to those of adult animals treated with saline. ∗∗P < 0.01; one-way ANOVA followed by the Bonferroni correction.
Credit
Bin Sun, Li Sun, Lixiang Zhang, Xinyu Xue, Qiuyan Tian, Lei Wu, Mei Li, Jian Huang, Hong Ni, Lixiao Xu, Chenxi Feng, Jing Ren, Hongliang Huo, Xia Zhang, Xing Feng, Wenhao Zhou, Wanliang Guo, Yaobo Liu, Rong Ju, Zhenlang Lin, Xiaofeng Yang, Xin Ding
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