RIP analyses were performed to detect the interaction between LncRNAHEIH/LncRNA-HULC and HBXIP using the Magna RIP RNA-Binding Protein Immunoprecipitation Kit (EMD Millipore Corporation, Billerica, MA, USA) in accordance with the manufacturer's instructions.
Total protein was extracted from the liver tissues and HepG2.2.15 cells, then quantified, diluted with 5x loading buffer to the same concentration, denatured at 95[degrees]C, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and then transferred onto polyvinylidene fluoride membranes, which were blocked with 5% skimmed milk at room temperature for 2 h and then incubated with Abs against HBXIP (dilution, 1 : 1000; Abcam) and GAPDH (dilution, 1 : 3000; Abcam) overnight at 4[degrees]C.
LncRNA-HEIH and LncRNA-HULC Coimmunoprecipitates with HBXIP. RIP assays were performed to determine whether HBXIP interacts with LncRNA-HEIH and LncRNA-HULC.
In the present study, the positive interactions of HBXIP with lncRNA-HULC and lncRNA-HEIH were further investigated by RIP analysis, which showed that HBXIP expression was greater in HBV-positive than HBV-negative HCC.
In summary, the present findings demonstrate that the interactions of lncRNA-HULC and lncRNA-HEIH with HBXIP might be involved in the occurrence of hepatitis B-related diseases.