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Fig. 7 | Acta Neuropathologica Communications

Fig. 7

From: Overexpression of Lin28A in neural progenitor cells in vivo does not lead to brain tumor formation but results in reduced spine density

Fig. 7

Phosphoproteomic analyses of GL mice. a-c MS-based phosphoproteomics of CTRL (n = 5) and GL cortices (n = 4) at P0. Hierarchical clustering of proteins showing significantly altered phosphosite abundance (decreased peptide phosphorylation levels in blue marked with dotted square) (a). Network analysis of GO Term Biological Processes based on decreased peptide phosphorylation levels displayed downregulated proteins of processes involved in microtubule dynamics (b). Network analysis of GO Term Molecular Functions and Cellular Component delivered the terms “dendritic shaft” and “protein kinase A binding” (c). d-f Detailed phosphosite analysis with upstream regulatory proteins and workflow (d). Decreased phosphosites were 82.35% serine sites, 5.88% tyrosine sites and 11.75% threonine sites (e). Venn diagram showing the top three upstream regulatory proteins RICTOR, RAPTOR and MPRIP regarding to phosphosite hits (f). g-p IHC staining of mTORC1 downstream signaling target Ser235/236 pS6 shows a stronger signal of positive cells in the cortex (l) and hippocampus (m-p) of GL mice when compared to CTRL mice (g, and h–k) at P15. q, r Western blot and quantification of pGSK3B, LIN28A, b-III TUBULIN, GSK3B, AKT and pAKT in cortices of GL and CTRL mice at P15 (every sample number represents an individual mouse). Significantly increased LIN28A (p = 0.0002, unpaired t-test, n = 5 for CTRL, n = 5 for GL) protein expression in GL mice compared to CTRL. No significant changes in pGKS3B/total GSK3B (p = 0.0766, unpaired t-test, n = 5 for CTRL, n = 5 for GL) and pAKT/total AKT ratios (p = 0.2270, unpaired t-test, n = 5 for CTRL, n = 5 for GL) in GL mice compared to CTRL (r). Scale bar in g is 100 µm for l; scale bar in insert of g is 25 µm for all inserts in g and l; scale bar in h is 250 µm for h–k, m-p. CA = cornu ammonis, DG = dentate gyrus

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