Fig. 3

a Schematic representation of the design for RNA-seq experiments; b plot showing DEGs from day 3 and day 6 control and Adnp−/− ESC-derived neurospheres; RNA-seq for each time points were repeated at least two times. DEGs were defined by FDR < 0.05 and a Log2 fold change > 1. The numbers of upregulated and downregulated genes were shown as average. c Heat map illustrating the expression of selected neuroectoderm and pluripotency-related genes that were shown as log2 FPKM in day 6 control and Adnp−/− ESC-derived neurospheres. Each lane corresponds to an independent biological RNA-seq sample. d Representative morphology of day 6 Adnp−/− ESC-derived neurospheres after addition of Tet-Express proteins from the indicated time points. Tet stands for Tet-Express. Experiments were repeated two times, and similar results were obtained. e IF staining of PAX6 for day 6 Adnp−/− ESC-derived neurospheres after addition of Tet-Express proteins at the indicated time points. Experiments were repeated three times, and similar results were obtained. f Quantification of mean fluorescence intensity of PAX6 staining using ImageJ for panel (e), based on three biologically independent experiments (n = 3–5 different regions of interest per group). Data are presented as mean values ± SEM and p values by two-tailed unpaired t-test are shown. ns not significant. Source data are provided as a Source Data file.