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. 2017 Apr 10;7:46204. doi: 10.1038/srep46204

Figure 5. LINC00305 associates with LIMR and AHRR.

Figure 5

(A) RNA-pull-down assay to identify LINC00305 binding proteins in THP-1 cells. The eluted proteins were separated by SDS-PAGE and subjected to silver staining (multiple exposures are presented in Supplementary Figure 6). Antisense RNA to LINC00305 (AS) was used as a negative control. The black arrow indicates the band representing the LINC00305-specific binding protein identified by mass spectrometry as LIMR. (B) RIP assay in HeLa cells transfected with LINC00305 and HA-tagged LIMR. RNA was immunoprecipitated using normal rabbit IgG or the anti-HA antibody. GAPDH was used as the negative control. **p < 0.01, ns: no significance vs. the indicated group. (C) GST pull-down assay to identify LIMR-interacting proteins in THP-1 cells. The eluted proteins were separated by SDS-PAGE and visualized using Coomassie Brilliant Blue Staining. The black arrow indicates the band representing the LIMR-specific interacting protein identified by mass spectrometry as AHRR. (D) Co-IP assay in 293 T cells transfected with LIMR and His-tagged AHRR. Proteins were immunoprecipitated using normal rabbit IgG or anti-His antibody. Input samples and the precipitated proteins were then analysed using anti-His and anti-LIMR antibodies (full length blots are presented in Supplementary Figure 9). (E) Co-IP assay in 293 T cells transfected with LIMR and His-tagged AHRR with or without LINC00305 (full length blots are presented in Supplementary Figure 10).