[Retracted] Novel lncRNA AL033381.2 Promotes Hepatocellular Carcinoma Progression by Upregulating PRKRA Expression

Figure 5

AL033381.2 interacts with PRKRA. (a, b) The specific biotin-labeled AL033381.2 probe was added to the cell lysate, and the precipitated proteins were resolved by SDS-PAGE and then stained with silver. The AL033381.2 probe was analyzed by using SDS-PAGE. The antisense of AL033381.2 and the beads serve as the control. The AL033381.2-specific bands (red rectangle) were excised and analyzed using mass spectrometry. (c) IPA analysis identified the genes located in the cytoplasm were associated with cell proliferation and apoptosis. (d) RIP assay showed that STAU1, PRDX2, ETFB, PRKRA, KRT8, and RNH1 interacted with AL033381.2 in Huh-7 cells by RT-qPCR analysis. (e) Western blot results exhibited the capability of PRKRA, PRDX2, RNH1, and ETFB binding to AL033381.2 in Huh-7 cells. (f) RNA pull-down assay was conducted in Huh-7 cells using biotinylated AL033381.2 or antisense RNA probe transcribed in vitro and measured by western blots. (g) RIP assay was employed in Huh-7 cells using an antibody against PRKRA to determine AL033381.2 RNA enrichment in immunoprecipitated complexes with IgG as a negative control. (h) Western blot analysis was applied to determine the protein level of PRKRA after transfection of the cells with shAL033381.2 or shCtrl.

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