Fig. 4. Rescue of cohesin-dependent gene expression.

a Schematic of NLS-TEV mediated RAD21-TEV degradation and rescue. b Western blot of RAD21-TEV protein expression over a time course of Dox treatment (6 h, 100 ng/ml). Bar plot of RAD21-TEV protein expression normalized to LAMIN B (n = 4). c Volcano plots of gene expression log₂ fold-change versus adjusted P value in RAD21-TEV + Dox treated neurons before and after rescue of RAD21-TEV expression (n = 3). Left: 570 genes were down- and 150 genes were upregulated following RAD21-TEV depletion (adj P < 0.05, Wald Test, Benjamini-Hochberg adjusted). Right: 43 genes were down- and 44 genes were upregulated after rescue of RAD21-TEV expression (adj P < 0.05, shown in red and orange, Wald Test, Benjamini-Hochberg adjusted). Genes in red are significantly deregulated in both RAD21-TEV depletion and rescue, genes in orange are significantly deregulated only after rescue (DE differentially expressed). d Log₂ fold-change of significantly deregulated genes (adj P < 0.05) following RAD21-TEV depletion (left) and after rescue (right). e The number of significantly deregulated genes (adj P < 0.05) after RAD21-TEV depletion (in green) and after rescue of RAD21-TEV expression (in orange). DE differentially expressed. f Heatmap of de novo deregulated genes following RAD21-TEV rescue (adj P < 0.05) and their maturation trajectory in control (left) and +Dox treated samples (right). g Barcode plot of de novo deregulated genes following RAD21-TEV rescue (adj P < 0.05) and their enrichment for directionality in maturation. Top: significantly de novo upregulated genes following rescue are enriched for genes upregulated during neuronal maturation. Bottom: de novo downregulated genes after cohesin rescue are enriched for genes that are downregulated during neuronal maturation.