Journal: PLOS Biology
Article Title: Sox5 controls the establishment of quiescence in neural stem cells during postnatal development
doi: 10.1371/journal.pbio.3002654
Figure Lengend Snippet: (A) Experimental approach of RNA-Seq analysis of FACS-sorted NSCs from Control and Sox5 null P14 mice. (B) Principal Component Analysis (PCA) of DEGs indicating independent segregation of the two genotypes analyzed, Control (gray) and Sox5 null (violet). Heat Map of DEGs in Sox5 null vs. Control. Volcano Plot of Log2FC and −log10 (padjusted) of DEGs between Control and Sox5 null mice. Several relevant genes are indicated in boxes, and those related to quiescence ( Foxo3, Foxo4, Huwe1 ) are downregulated, whereas genes related to cell cycle ( Myc ) and BMP canonical signaling ( BMP2, BMP4, BMP5 ) are upregulated in Sox5 null vs. Control DG. (C) Enrichment analysis of GO terms related to Biological Processes and Molecular Functions in DEGs of Sox5 null vs. Control NSCs. (D) KEGG and PANTHER pathways analysis of DEGs of Sox5 null vs. Control NSCs. DEGs were selected for their p -adjusted value < 0.05. (E, F) Node graphs of most representative upregulated cell cycle genes and downregulated TGF-β/BMP Superfamily signaling pathway genes generated by the PANEV software (PAthway NEtwork Visualizer; v. 17.0). (G) Heat Map of DEGs manually curated in the list of BMP signaling pathway genes comparing Sox5 null vs. Control NSCs. (H) Quantitation of the relative levels of mRNA expression by quantitative PCR for the indicated transcripts in Sox5 null vs. Control NSCs prepared in vitro (see ). Results are shown as 2 −ΔΔCT normalized with respect to GAPDH mRNA and relative to P14 NSCs Control values (dashed line y-axis = 1). Control and Sox5 null P14 mice ( N = 3 and 3, respectively) were used to prepared DG NSC cultures and were analyzed for each condition and experimental replicates were performed. (I) Scheme of the BMP canonical signaling pathway. Data represent mean values ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001according to a Student t test. Data are available in as a part of Supporting information. Drawings were created through SciDraw, Bioicons, Bioart, and Inkscape.
Article Snippet: The following Taqman assays probes were used: Anapc (Mm00614339_m1), Axin2 (Mm00443610_m1), Bmp2 (Mm01340178_m1), Bmp3 (Mm00557790_m1), Bmp7 (Mm00432102_m1), Bmpr1b (Mm03023971_m1), Ccna2 (Mm00438063_m1), Ccnb1 (Mm03053893_gH), Ctdsp1 (Mm00778482_s1), Ctdsp2 (Mm01254394_m1), Fstl5 (Mm00618418_m1), Gapdh (Mm99999915_g1), Huwe1 (Mm00615533_m1), Id4 (Mm00499701_m1), Mcm4 (Mm00725863_s1), Myc (Mm00487804_m1), Ppm1a (Mm00725963_s1), Ppm1h (Mm00620945_m1), Smurf2 (Mm03024086_m1), Sox5 (Mm01264584_m1) and Ythdf2 (Mm00661925_m1).
Techniques: RNA Sequencing, Control, Generated, Software, Quantitation Assay, Expressing, Real-time Polymerase Chain Reaction, In Vitro