Supplementary Figure 2: Knockdown of cytoplasmic lnc-SCA7.
From: Cross-talking noncoding RNAs contribute to cell-specific neurodegeneration in SCA7
(A) Sequence alignment between regions in mouse Atxn7l3 and lnc-SCA7 used to design specific short hairpin RNAs (shRNAs) against lnc-SCA7. The initial base of the predicted binding sites in these transcripts is noted to the left of the alignment. Identical nucleotides between Atxn7l3 and lnc-SCA7 are denoted by vertical lines. Regions targeted by the siRNAs are highlighted in blue. (B) Expression levels of lnc-SCA7 (blue) and Atxn7 (red) following knockdown of lnc-SCA7 in N2A cells using each of the 3 designed shRNAs (sh-lnc-SCA7, sh-lnc-SCA7_1, and sh-lnc-SCA7_2). (C) lnc-SCA7 (blue) and Atxn7 mRNA (red) are found predominantly in the cytoplasm of N2A cells (y-axis). Malat1 (light grey), a known nuclear transcript, was used as control. Fold enrichments (expression level measured in the cytoplasmic relative to the expression level measured in the nucleus) for all genes were measured and normalized against that of Gapdh, a cytoplasmic single exonic transcript. Error bars represent s.d.m. n = 3 cell cultures per condition; * p < 0.05; ** p < 0.01; Two-tailed Student’s t-test.
