Figure 2.

TAT-BIND Suppressed GGGGCC RNA Foci Formation and RAN Translation in pAg3-(GGGGCC)₆₆-Expressing SK-N-MC Cells

(A) TAT-BIND suppressed GGGGCC RNA foci formation in pAg3-(GGGGCC)₆₆-expressing SK-N-MC cells. In situ hybridization was performed to detect GGGGCC RNA foci (red) by using a TYE563-labeled LNA probe. Nuclei (blue) were stained by Hoechst 33332. The scale bar represents 10 μm. (B) Quantification of the percentage of cells containing RNA foci after transfection. (C) TAT-BIND suppressed poly-GR protein expression in pAg3-(GGGGCC)₆₆-expressing cells. (D) Statistical analysis of band intensity (poly-GR/GAPDH) of (C). (E) TAT-BIND suppressed poly-GA protein expression in pAg3-(GGGGCC)₆₆-expressing cells. (F) Statistical analysis of band intensity (poly-GA/GAPDH) of (E). (G) TAT-BIND treatment suppressed poly-GP protein expression in pAg3-(GGGGCC)₆₆-expressing cells. (H) Statistical analysis of band intensity (poly-GP/GAPDH) of (G). 1 μg pAg3-(GGGGCC)₆₆ plasmid was used to transfect SK-N-MC cells, followed by application of 10 μM of each peptide. Cells were collected and lysed for western blot analysis 48 h after treatment. GAPDH was used as the loading control. Only representative blots are shown. All experiments were repeated at least thrice with consistent results. Data were plotted as mean ± SEM. ***p < 0.001 and ****p < 0.0001; NS, no significance.