Figure 1. Sam68 is highly expressed in NPCs and decreases during differentiation.
(A) qPCR analysis of Khdrbs1 mRNA levels in the cortex of embryonic (E10.5-E15.5) and post-natal (P0-25dpp) mouse brain. Khdrbs1 relative expression was evaluated by △CT method using L34 expression for normalization. (B) Western blot analysis of Sam68 and SOX2 expression in lysates from embryonic (E10.5-E15.5) and post-natal (P0-25dpp) mouse cortices. GAPDH was used as loading control. (C and D) Immunofluorescence analyses of Sam68 and SOX2 expression in E13.5 mouse brain. (C) Horizontal sections of whole brain; white arrows point to periventricular zones where both proteins are highly expressed. Scale bar = 250 µm. (D) High-magnification confocal images confirm Sam68 and SOX2 colocalization in most cells of the VZ and SVZ. Scale bar = 25 µm. (E) High magnification of confocal images of 1 dpp mouse VZ-SVZ, show the colocalization of Sam68 and SOX2 in NPCs (white arrows). Scale bar = 25 µM. (F) Analysis of Sam68, SOX2 and TUBB3 mRNA (left panels) and protein levels (right panels) in NPCs cultured under proliferating condition (0d) or during 1–6 days of differentiation (1-6d). (G) qPCR analysis of Khdrbs1, Sox2 and Tubb3 mRNA levels in NPCs under proliferation conditions (0d) and 1–6 days of differentiation (1, 3, 6d). Relative expression was evaluated using △△CT method and 0d as reference point. L34 expression was used for the initial △CT normalization. NPCs, Neural progenitor cells.