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. 2017 May 3;15(5):e2001220. doi: 10.1371/journal.pbio.2001220

Fig 1. Suppressor of Mek null (Smek) expression and function in mouse cortical development.

Fig 1

(A) Fixed cyroembedded coronal sections from E12.5 or E14.5 mouse forebrain stained with antibodies against Tuj1 (red), Tbr1 (green), and microtubule-associated protein 2 (MAP2) (red). Nuclear staining is shown by 4',6-diamidino-2-phenylindole (DAPI) (blue). (Right) Quantification of Fig 1A and S1D Fig. (E12.5: WT, n = 4, dKO, n = 5; E14.5: WT, n = 6, dKO, n = 5). Scale bars, 50 μm. (B) Same as Fig 1A for Pax6. (Right) Quantification of Fig 1B (E12.5: WT, n = 5, dKO, n = 4; E14.5: WT, n = 6, dKO, n = 3). Scale bar, 50 μm. (C) Immunostaining with Nestin (green) and Tuj1 (red) antibodies in WT and Smek1/2 dKO neural progenitor cells (NPCs). Nuclear staining is shown by DAPI (blue). Scale bars, 50 μm. (Lower) Quantification of anti-Tuj1–positive (WT-Un, n = 3; WT-2DIV, n = 6; dKO-Un, n = 3; dKO-2DIV, n = 6) and anti-Nestin–postive (WT-Un, n = 6; WT-2DIV, n = 6; dKO-Un, n = 6; dKO-2DIV, n = 6) cells in Fig 1C. Un, undifferentiation; 2Diff, differentiation 2 d in vitro (DIV). (D) Single cells of WT and Smek1/2 dKO NPCs were separated by serial dilution and sphere formation was induced for 8 d in vitro. Relative size of primary spheres grown up to 8 DIV were quantified by the ImageJ quantification software. Scale bars, blue (100 pixel), red (200 pixel). (E) Quantitative PCR (qPCR) analysis of indicated mRNAs. Values correspond to the average ± SD. Diff. (d), days in differentiation. Statistical t test analysis was performed to calculate significance (*p < 0.05, **p < 0.005, ***p < 0.0005; not significant (ns), p > 0.05). All quantification data underlying panels A–D can be found in S2 Data.