Research Paper Volume 11, Issue 17 pp 6657—6673

Figure 4. MIR4435-2HG binds to DSP protein and reduces DSP expression. (A) RNA pull-down experiments were performed to identify proteins interacting with MIR4435-2HG. Silver staining revealed a specific band associated with biotinylated sense MIR4435-2HG. (B) The biotinylated sense MIR4435-2HG band was excised, digested, and subjected to mass spectrometry, which identified DSP and HRNR as the MIR4435-2HG-interacting proteins. (C) Levels of DSP and HRNR were confirmed in the shMIR4435-2HG (HGC-27 cells) and control groups using RT-qPCR (**P < 0.01). (D) The interaction of DSP with MIR4435-2HG or HRNR with MIR4435-2HG by RNA pull-down assay. DSP was detected within sense MIR4435-2HG pull-down protein complexes but not with antisense MIR4435-2HG. (E) Expression levels of DSP within HGC-27/shMIR4435-2HG-xenografted and HGC-27/shNC-xenografted tumors were detected using WB and IHC (**P < 0.01).