Lineage-specific decline of E2F1 nuclear levels during oligodendrocyte differentiation. Lineage-specific decline of E2F1 nuclear levels during oligodendrocyte.

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Lineage-specific decline of E2F1 nuclear levels during oligodendrocyte differentiation. Lineage-specific decline of E2F1 nuclear levels during oligodendrocyte differentiation. A, Transcript levels of Pdgfrα, Cnpase, Mag, Mbp, Plp, Mog in rat OPCs cultured for 5 d in the presence of mitogens (PDGF + FGF2) or in conditions favoring differentiation into oligodendrocytes (T3) or astrocytes (BMP4) were analyzed by quantitative RT-PCR. B, Transcript levels of E2fs (E2f1, E2f3, E2f4, E2f5) were analyzed by quantitative RT-PCR in the same samples described in A. C, E2f1 transcript level in RNA samples isolated from OPC either kept in the presence of mitogens (D0), or differentiated for 1, 3, or 5 d. Transcript levels were normalized by the levels of Gapdh and shown as relative to the levels detected at D0. Bar graphs represent the average values and error bars represent SD (n = 3, *p < 0.05 **p < 0.01, ***p < 0.005). D, Protein expression and subcellular localization of E2F1 during rat OPC differentiation in cultured cells. OPCs were stained for E2F1 (red) and for the OL marker A2B5 (green in D0) or O4 (green in D1 and D3). Note the nuclear localization of E2F1 at D0 followed by its cytoplasmic localization as cells exited the cell cycle (D1). E, Bar graphs denote the percentage of cells with nuclear or cytosolic localization of E2F1+ during rat OPC differentiation at the indicated time points. **p < 0.005, two-way ANOVA. Scale bar in D, 20 μm. Laura Magri et al. J. Neurosci. 2014;34:1481-1493 ©2014 by Society for Neuroscience