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. 2008 Nov 5;28(45):11720–11730. doi: 10.1523/JNEUROSCI.1932-08.2008

Figure 5.

Figure 5.

miR-9 downregulates PMP22 in Schwann cells. A, miR-9 reduces PMP22 levels in Schwann cells. The Schwann cells transiently transfected with miR-9 contain reduced steady-state levels of PMP22 mRNA (*p < 0.05, Student's t test, versus miR-Neg, transfection with the empty plasmid, n = 3). Lipo, Lipofectamine negative control. The 18S ribosomal RNA is used for normalization. Error bars represent SD. B, Western blot analysis of Schwann cells after miR-9 transfection. The quantification of the downregulation of PMP22 protein after transfection with miR-9 is obtained from three independent experiments (*p < 0.05, Student's t test, versus miR-Neg, n = 3). GAPDH is used for normalization and N-glycosidase (PNGase) treated cell lysate is shown as a control for PMP22. The PMP22 specific bands are shown by arrows. NS, Nonspecific band; Lipo, lipofectamine negative control. Error bars represent SD. C, Immunofluorescence analysis of Schwann cells after miR-9 transfection. A reduction of PMP22 immunoreactivity is seen after transfection of Schwann cells with the pcDNA6.2-GW/EmGFP-miR-9 plasmid (e, shown by arrow), compared with the cells transiently transfected with the miR-Neg plasmid (b, shown by arrow). The transfected Schwann cells are detected by EmGFP autofluorescence (a–f, arrows). Nuclei are visualized by Hoechst staining. Scale bar, 10 μm.