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. 2011 Mar 7;192(5):797–811. doi: 10.1083/jcb.201007014

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© 2011 Laursen et al.

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Figure 6. — The subcellular distribution of hnRNP-K changes during oligodendrocyte differentiation and myelination. (A) The top panels (labeled DAY 1) show OPCs differentiated for 1 d and stained for hnRNP-K (green) and NG2 (red). Note that hnRNP-K is found almost entirely in the nucleus (top). The bottom panels (labeled DAY 4) show oligodendrocytes generated by differentiation for 4 d and stained for hnRNP-K (green) and MBP (red). Note that more hnRNP-K is found outside the nucleus as compared with d 1, and that some is localized to the myelin sheets. (B) hnRNP-K localization to different cellular compartments (Cytopl, cytoplasmic; Membr, membrane; Nucle, nuclear; Cytosk, cytoskeleton) analyzed by fractioning of OPCs or oligodendrocytes generated by differentiation for 4 d (D4) followed by Western blotting. (C) Quantification of Western blots from the cell fractioning assays shown in B. The numbers are mean values ±SD of three independent experiments, with statistical significance tested by two-way ANOVA followed by a Boniferroni post-test. *, P < 0.05. Note the shift of hnRNP-K from nuclear to membrane fraction upon differentiation. (D) Oligodendrocytes and DRG neurons co-cultured for 12 d and stained for hnRNP-K (green), MBP (red, to identify differentiated oligodendrocytes), NG2 (purple, to identify undifferentiated OPCs), and neurofilament (blue, to identify axons) as indicated. Note the strong nuclear staining for hnRNP-K for NG2+ OPCs (arrowheads), the translocation of hnRNP-K to the internodes in the MBP+ oligodendrocytes (arrows) shown in the top row of images, and the absence of nuclear hnRNP-K in some MBP+ cells (black and white arrows) shown in the bottom row of images. (E) Oligodendrocytes and DRG neurons co-cultured for 12 d and stained for hnRNP-K (green), β1-integrin (red), MBP (purple), and neurofilament (blue) as indicated. The smaller panel on the right shows a high power image of the internodes highlighted by a box in the middle panel. Note the hnRNP-K staining and colocalization with β1-integrin along these internodes.

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