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. 2020 Dec 8;15(6):1317–1332. doi: 10.1016/j.stemcr.2020.11.006

Figure 2.

Figure 2

MeCP2 Has a Role in MajSat Forward Transcript Targeting to PCH

(A) Top: Strand-specific RT-qPCR analysis of MajSat-fw and MajSat-rv transcripts in mESC-derived TK23_WT and Mecp2−/y neurons. Data are normalized to Gapdh and are means ± SD of transcript levels relative to TK23_WT, from three independent experiments. NS, not significant (two-tailed Student's t test). Bottom: Electrophoretic analyses of endpoint strand-specific RT-qPCR for the MajSat-fw and MajSat-rv RNAs. RT, reverse transcriptase; nc, negative control. (B, C) Left: Representative images of i3D-RNA FISH using the MajSat-fw LNA probe in TK23_WT and Mecp2−/y undifferentiated cells (B) and mESC-derived neurons (C) without and with RNase A treatment. Chromocenters were stained with DAPI. Scale bar, 25 μm. Right: Quantification of MajSat-fw RNA signals in the three-dimensional space of each nucleus. Data are shown as box and whisker plots with 200 nuclei analyzed per condition, from two replicate slides. ∗∗∗p < 0.001 (two-sample Kolmogorov-Smirnov test). See also Figure S1.