Supplementary Figure 6: 15.5K–U8 snoRNA EMSA control experiments. | Nature Genetics

Supplementary Figure 6: 15.5K–U8 snoRNA EMSA control experiments.

From: Mutations in SNORD118 cause the cerebral microangiopathy leukoencephalopathy with calcifications and cysts

Supplementary Figure 6

(a) Addition of an antibody to 6×His (ab18184) causes a supershift, demonstrating that the observed shift in U8 RNA is a result of binding to His-15.5K. (b) A competition assay to demonstrate binding specificity. An excess of unlabeled in vitro–transcribed wild-type U8 RNA (2.5 μg) is sufficient to outcompete 5′-end-labeled in vitro–transcribed U8 RNA and eliminate the band shift. (c) Coomassie-stained gel showing purified His-15.5K protein. Lane 1, Precision Plus Protein Dual-Color Standard (Bio-Rad); lane 2, purified His-15.5K protein.

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