Fig. 6: Characterization of the defects in heterochromatin anchoring and the formation of type I NB lineages induced by KD of tsh or Arm in type I NBs. | Experimental & Molecular Medicine

Fig. 6: Characterization of the defects in heterochromatin anchoring and the formation of type I NB lineages induced by KD of tsh or Arm in type I NBs.

From: Baf-mediated transcriptional regulation of teashirt is essential for the development of neural progenitor cell lineages

Fig. 6

a Representative images of immunostaining for Dpn (teal) and Pros (magenta) in control type I NBs and type I NBs expressing Baf RNAi (Baf KD), tsh RNAi (tsh KD), and Arm RNAi (Arm KD) in brains of third-instar larvae (ase-Gal4/+; UAS-mCD8GFP/+, ase-Gal4/+;UAS-Baf RNAi/UAS-mCD8GFP, ase-Gal4/+;UAS-tsh RNAi/UAS-mCD8GFP, and ase-Gal4/+;UAS-Arm RNAi/UAS-mCD8GFP). The white dashed lines indicate the outlines of NBs (second row) and neural progenies (third row) (Scale bar, 20 μm). b Representative images of immunostaining for H3K9me3 (magenta) with Hoechst staining in cultured primary control type I NBs and type I NBs expressing Baf RNAi (Baf KD), tsh RNAi (tsh KD), and Arm RNAi (Arm KD) that were isolated from the brains of third-instar larvae (ase-Gal4/+; UAS-mCD8GFP/+, ase-Gal4/+;UAS-Baf RNAi/UAS-mCD8GFP, ase-Gal4/+;UAS-tsh RNAi/UAS-mCD8GFP, and ase-Gal4/+;UAS-Arm RNAi/UAS-mCD8GFP). NBs were visualized by expression of the fluorescent plasma membrane marker protein mCD8GFP. The white dashed lines in the panels show the outlines of the inner nuclear membrane (Scale bar, 10 μm). c Stacked bar graph showing the proportion of mean intensity of heterochromatin localized in three zones of cultured primary type I NBs expressing the transgenes indicated in (b). The number of type I NBs tested was as follows: Control = 6, Baf KD = 7, tsh KD = 11, and Arm KD = 5. d Scatter dot plots displaying the proportion of mean intensity of heterochromatin localized in the peripheral zone of type I NBs expressing the transgenes indicated in (b). ***P < 1.0 × 10−3; ****P < 1.0 × 10−4 by one-way ANOVA with Tukey’s post hoc test; error bars, mean ± SEM. The number of type I NBs tested was as follows: Control = 6, Baf KD = 7, tsh KD = 11, and Arm KD = 5. e Scatter dot plots displaying the proportion of mean intensity of heterochromatin localized in the central zone of type I NBs expressing the transgenes indicated in (b). **P < 1.0 × 10−2; ***P < 1.0 × 10−3 by one-way ANOVA with Tukey’s post hoc test; error bars, mean ± SEM. The number of type I NBs tested was as follows: Control = 6, Baf KD = 7, tsh KD = 11, and Arm KD = 5. f Representative images of immunostaining for Baf (magenta) and tsh (teal) in cultured primary type I NBs that were isolated from the brains of third-instar larvae (ase-Gal4/+; UAS-mCD8GFP/+). NBs are visualized by expression of the fluorescent plasma membrane marker protein mCD8GFP. The white dashed lines in the panels show the outlines of the inner nuclear membrane (Scale bar, 10 μm).

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