Fig. 6: DLC1 determines the increased vulnerability of trunk NCCs to PB treatment. | Nature Communications

Fig. 6: DLC1 determines the increased vulnerability of trunk NCCs to PB treatment.

From: Determination of trunk neural crest cell fate and susceptibility to splicing perturbation by the DLC1-SF3B1-PHF5A splicing complex

Fig. 6

a Experimental strategy. b Ectopic DLC1 expression in somites reduced expression of SOX9 (b, c) and SNAI2 (d, e) in a cell-autonomous manner (open arrowheads) upon PB treatment. Open arrowheads indicate loss of SOX9 and SNAI2 expression in pre-migratory NCCs. There was no alteration of SOX9 (b, c) and SNAI2 (d, e) expression in DLC1-overexpressing somites and in NCCs (black and white arrows) in the PBS treatment group. Scale bar, 50 µm. Yellow dotted boxes outline the magnified view in the rightmost column of each panel. n = 5 embryos per treatment. f IP shows the association of ectopic DLC1 with endogenous PHF5A and SF3B1 in somites of embryos treated with PBS Ctrl and PB. g, i Electroporation of DLC1 in somites resulted in increased levels of SOX9 intron 2 and SNAI2 intron 1 in the presence of PB compared to PBS Ctrl. n = 5 embryos per treatment. h, j Schematic diagrams depict the presence of intron 2-retained SOX9 pre-mRNA (h) and intron 1-retained SNAI2 pre-mRNA (j) in DLC1 overexpressing somites of embryos treated with PB. k The HCR analysis revealed the expression of SOX9 intron 2 and SNAI2 intron 1 in DLC1-overexpressing somites (So) of embryos treated with PB, as compared to the PBS Ctrl. l–o Quantification of fluorescence intensities. 20 sections per treatment were used for the quanitification. Scale bar: 50 µm. The minima, maxima, center, percentile values and the exact p-values are listed in the Source data file. Mean ± SD. ns: no significant difference, ****P < 0.0001. NT neural tube, So somites.

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