Temporal Analysis of Genome-wide Transcription Changes in Astrocyte Reprogramming
(A) Schematic representation of the experimental procedure inducing the activation of Neurog2ERT2-IRES-DsRed or Ascl1ERT2-IRES-DsRed by tamoxifen (OHT indicated by uppermost black bars) for reprogramming astrocytes into neurons.
(B, C, E, and F) Micrographs of astrocytes infected with the constructs indicated in red on the left side and immunostained for the astrocytic marker GFAP (green) and the neuronal marker βIII-tubulin (white). Scale bars, 100 μm.
(D and G) Quantification of non-reprogrammed cells (GFAP) or reprogrammed cells (βIII-tubulin) without or with OHT 8 days post-induction (DPI). Mean ± SEM; n = 4 independent experiments; statistical test: two-tailed Mann-Whitney test (∗p < 0.05).
(H) Schematic representation of the experimental procedure for genome-wide mRNA analysis.
(I) Heatmap of genes regulated by both Neurog2ERT2 and Ascl1ERT2 within 24 hr after induction by OHT.
(J) Venn diagram of genes regulated by Neurog2ERT2 or Ascl1ERT2 24 hr after OHT.
(K and L) Real-time qPCR) analysis on selected candidates upon Neurog2ERT2 (K) or Ascl1ERT2 (L) induction by OHT for 24 hr. Mean ± SEM; n = 3 independent experiments.
See also Figure S1, Table S1, Table S2, Table S3, Table S4, and Table S5.