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. 2022 Jan 4;11:e72195. doi: 10.7554/eLife.72195

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© 2022, Guo et al

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Figure 4. — (A) Schematic of experimental workflow to biochemically isolate and sequence translated mRNAs from Gli1+ RGLs (Gli1^CreERT2:Rpl22HA^f/+:Klf9^{f/f or +/+} mice). n = 3 mice, 6 dentate gyri/sample, 3 samples per group. (B) Principal component analysis (PCA) plot of translational profiles of Gli1^CreERT2-targeted Klf9^{+/+ or f/f} RGLs. First two principal components are shown with the corresponding fractions of variance. (C) Left: heatmap of expression values for differentially expressed genes. Middle: volcano plot of statistical significance (−log10 p value) vs. magnitude of change (log2 of fold change) of gene expression. Differentially expressed genes are marked in red. Upregulated genes in Klf9^f/f RGLs are on the right and downregulated genes in Klf9^f/f RGLs are on the left. Right: pie chart of numbers of upregulated and downregulated genes in Gli1^CreERT2-targeted Klf9^f/f RGLs. (D) qRT-PCR on biochemically isolated mRNAs from Gli1^CreERT2:Rpl22HA^f/+:Klf9^{f/f or +/+} mice validating candidate differentially expressed genes. n = 3 samples, 6 dentate gyri/sample, 3 samples per group. (E) Immunostaining and quantification of Notch1 intracellular domain (NICD) in RGLs of Gli1^CreERT2: Klf9^{f/f or +/+} mice. Deletion of Klf9 results in increased NICD levels in RGLs consistent with Lnfg-dependent potentiation of Notch1 signaling (Cartoon, top left). n = 3 mice/group. Data are represented as mean ± standard error of the mean (SEM). *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar: 10 μm.

Figure 4—figure supplement 1. — (A) Schematic of experimental workflow to biochemically isolate and sequence translated mRNAs from Gli1+ RGLs (Gli1^CreERT2:Rpl22HA^f/+:Klf9^{f/f or +/+} mice). n = 3 mice, 6 dentate gyri/sample, 3 samples per group. (B) Principal component analysis (PCA) plot of translational profiles of Gli1^CreERT2-targeted Klf9^{+/+ or f/f} RGLs. First two principal components are shown with the corresponding fractions of variance. (C) Left: heatmap of expression values for differentially expressed genes. Middle: volcano plot of statistical significance (−log10 p value) vs. magnitude of change (log2 of fold change) of gene expression. Differentially expressed genes are marked in red. Upregulated genes in Klf9^f/f RGLs are on the right and downregulated genes in Klf9^f/f RGLs are on the left. Right: pie chart of numbers of upregulated and downregulated genes in Gli1^CreERT2-targeted Klf9^f/f RGLs. (D) qRT-PCR on biochemically isolated mRNAs from Gli1^CreERT2:Rpl22HA^f/+:Klf9^{f/f or +/+} mice validating candidate differentially expressed genes. n = 3 samples, 6 dentate gyri/sample, 3 samples per group. (E) Immunostaining and quantification of Notch1 intracellular domain (NICD) in RGLs of Gli1^CreERT2: Klf9^{f/f or +/+} mice. Deletion of Klf9 results in increased NICD levels in RGLs consistent with Lnfg-dependent potentiation of Notch1 signaling (Cartoon, top left). n = 3 mice/group. Data are represented as mean ± standard error of the mean (SEM). *p < 0.05, **p < 0.01, ***p < 0.001. Scale bar: 10 μm.