Fig. 3: TGFβ-mediated enhancer-cluster assembly depends on JMJD3.

a UCSC Genome Browser captures show 4C-seq profiles spanning 200 kb around the VP enhancer (black arrow) in NSCs untreated or treated (3 h) with TGFβ. ChIP-seq signals of SMAD3 and JMJD3 upon TGFβ stimulation (0.5 and 3 h, respectively) are shown. The location of the members of the EC is also indicated with yellow boxes. b Capture showing a zoom into a region where TGFβ-induced contacts are lost in JMJD3-depleted (shJMJD3) NSCs. c Boxplot displays the averaged values obtained from two biological independent replicates of RPM signals of the peaks located 500 kb around the VP - excluding the nearest ±20 kb - (mm10 chr7:33841896-35860773) in control or shJMJD3 NSCs untreated or treated with TGFβ during 3 h. An independent region located in another chromosome (mm10 chr4:33076383-35216108) was used as a negative control. n = 2 biologically independent replicates were quantified. p-values are the result of a Wilcoxon-Mann–Whitney test. d Control NSCs or shJMJD3 NSCs were treated for 3 h with TGFβ. Then, total RNA was prepared, and the levels of the mRNA of the indicated genes were determined by qPCR. Values were normalized to the housekeeping gene Gapdh. The figure shows values relative to time 0 h. Results are the mean of three biologically independent experiments. Data are presented as mean values +/− SEM. **p < 0.01; **p < 0.001 (P values were calculated using two-tailed Student’s t test, p = p = 0.001131677 and p = 0.003848794). Source data are provided as a Source Data file.