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. 2015 Aug 5;290(37):22485–22493. doi: 10.1074/jbc.M115.658104

FIGURE 1.

FIGURE 1.

The MeCP2 NLS is critical for nuclear localization, and MeCP2-R270X localizes exclusively to the nucleus. A, schematic of MeCP2 fusion constructs with GFP and RFP used in localization experiments. MBD, methyl-CpG binding domain (amino acids 78–162); TRD transcriptional repression domain (amino acids 207–310), NLS, the NLS (amino acids 255–271) as described in Ref. 4. The schematic is not to scale. B, N2a cells were transiently transfected with MeCP2 tagged on the C terminus with GFP and live-imaged using endogenous GFP excitation with Hoechst 33342 as a nuclear counterstain. Individual channels are indicated, and a merged image overlaid on the differential interference contrast (DIC) channel is shown to indicate nuclear and cytoplasmic boundaries. The MeCP2 variants used are indicated. ΔNLS, full-length MeCP2 with amino acids 255–271 deleted. Scale bars = 10 μm. n = 3. C, a similar experiment as described in B except that N2a cells were transfected with MeCP2 tagged at its N terminus with RFP. Scale bars = 10 μm. n = 3.