Fig. 7: BAF53B rescues the differentiation defect in JMJD3/UTX-deficient human NPCs.

a Upper panel, qRT-PCR and western blot analysis of BRG1 and BRM in the indicated ESCs, NPCs, and their differentiated cells at day 28. Bottom panel, western blot analysis of BRG1 and BRM in the indicated differentiated wild-type (WT) or dKO cells at day 28 with the proteasome inhibitor MG132. The data represent the mean ± SD (standard deviation) from three independent replicates (n = 3). npBAF, neural progenitor cell-specific BAF complex. nBAF, neuron-specific BAF complex. dKO, deletion of both JMJD3 and UTX. b qRT-PCR and immuno-staining analysis of the npBAF component BAF53A in the indicated ESCs, NPCs, and their differentiated cells at day 28. The significance level was determined using unpaired two-tailed Student’s t-tests. **P < 0.01. The data represent the mean ± SD from three independent replicates (n = 3). c qRT-PCR and immuno-staining analysis of the nBAF component BAF53B in the indicated ESCs, NPCs, and their differentiated cells at day 28. The significance level was determined using unpaired two-tailed Student’s t-tests. *, P < 0.05. The data represent the mean ± SD from three independent replicates (n = 3). d Genomic views of the H3K27me3 modification and UTX and JMJD3 binding of BAF genes in the indicated cell lines. e Morphological examination and immuno-staining for the NPC markers SOX2/NES, the neuronal/glial markers MAP2/GFAP in dKO NPCs (H1-dKO + BAF53B) with forced BAF53B expression during random differentiation at day 1 and day 28. The numbers of MAP2⁺ or GFAP⁺ cells were analysed. Significance was determined by unpaired two-tailed Student’s t-tests. **P < 0.01. The data represent the mean ± SD from three independent replicates (n = 3). Scale bar, 50 μm. f Western blotting for H3K27me3 or BAF53B in the indicated wild-type, dKO, or BAF53B-rescued dKO differentiated cells at day 28. g Signal densities for OC or CO regions in the indicated wild-type (WT), dKO, or BAF53B-rescued dKO differentiated cells at day 28 from ATAC-seq data. OC, open-to-closed, indicates regions in dKO NPCs with reduced chromatin accessibility. CO, closed-to-open, indicates regions in dKO NPCs with increased chromatin accessibility. Error bars throughout the figure represent the SD (standard deviation) from three independent replicates (n = 3). Source data are provided as a Source Data file. More information is also provided in Supplementary Fig. 5. NS no significance.