Fig. 5: Non-brain-like isoforms alter migration and proliferation of cortical NSCs. | Nature Communications

Fig. 5: Non-brain-like isoforms alter migration and proliferation of cortical NSCs.

From: ZMYND11 functions in bimodal regulation of latent genes and brain-like splicing to safeguard corticogenesis

Fig. 5

a Schematic outlining functional assays after specific isoform knockdown in long-term neural stem cells (LTNSCs). b Migration assays (0 h, 24 h, 48 h) on LTNSCs with different isoform knockdown and quantification of scratch coverage compared with shNT (n = 48 areas across 3 independent experiments). Dashed lines mark migration. Red marks significantly changed shRNA. Boxplot shows median, quartiles, and whiskers (min/max). NT non targeting. c BrdU-PI assays on LTNSCs with MEAF6 long (brain) or short (non-brain) isoform knockdown with quantification (mean ± SEM, n = 3). d Schematic outlining splicing factor screening and semi-quantitative PCR on CTTN exon 11 and MEAF6 exon 6 (n = 2). Red rectangles mark ZMYND11 shRNA, blue rectangles mark significantly changed shRNA. Percent non-brain isoform values shown below the gels. e Heatmap of non-brain isoform percent values (row Z-score transformation) for (d) using selected high-confidence DAS events matching non-brain tissues and ZMYND11 KO. APP events are combined due to proximity. Statistics in (b): one-way ANOVA followed by Dunnett’s test. Statistics in (c): one-way ANOVA followed by Tukey’s test. ns P  >  0.05, *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. P values in (b): 24 h: shL1CAM_e3-L vs shNT, P < 0.0001; shL1CAM_e3-S vs shNT, P = 0.9996; shL1CAM_e28-L vs shNT, P = 0.0087; shL1CAM_e28-S vs shNT, P = 0.3781; shCTTN-L vs shNT, P = 0.2084; shCTTN-S vs shNT, P = 0.9971; shMEAF6-L vs shNT, P = 0.9786; shMEAF6-S vs shNT, P < 0.0001; 48 h: shL1CAM_e3-L vs shNT, P < 0.0001; shL1CAM_e3-S vs shNT, P = 0.9996; shL1CAM_e28-L vs shNT, P = 0.0151; shL1CAM_e28-S vs shNT, P = 0.4839; shCTTN-L vs shNT, P = 0.6008; shCTTN-S vs shNT, P = 0.9888; shMEAF6-L vs shNT, P = 0.9575; shMEAF6-S vs shNT, P < 0.0001. P values in (c): shMEAF6-L vs shNT for G1%, P = 0.1074; shMEAF6-S vs shNT for G1%, P = 0.0012; shMEAF6-L vs shMEAF6-S for G1%, P = 0.0002; shMEAF6-L vs shNT for S%, P = 0.0543; shMEAF6-S vs shNT for S%, P = 0.0002; shMEAF6-L vs shMEAF6-S for S%, P < 0.0001; shMEAF6-L vs shNT for G2%, P = 0.3799; shMEAF6-S vs shNT for G2%, P = 0.0055; shMEAF6-L vs shMEAF6-S for G2%, P = 0.0015. Source data are provided as a Source Data file.

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