Research Paper Volume 15, Issue 21 pp 12476—12496
lncRNA HOXC-AS2 promotes the progression of hypopharyngeal cancer by binding to the P62 protein mediating the autophagy process
Figure 4. HOXC-AS2 can bind to the SQSTM1 (P62) protein. (A) Analysis of the RNA pulldown results by silver staining after HOXC-AS2 overexpression. (B) Bubble plots showing the results of the pathway analysis. (C) GO enrichment analysis of proteins identified in the HOXC-AS2 RNA pulldown assay. The P62 protein was found to be closely related to autophagy. (D) RIP assay of HOXC-AS2 with SQSTM1 and PHB2 proteins. (E) The mRNA expression level of P62 was confirmed through qRT-PCR analysis after HOXC-AS2 overexpression. (F) The mRNA expression level of P62 was confirmed through qRT-PCR analysis after HOXC-AS2 knockdown. (G) HOXC-AS2 overexpression plasmids were transfected into FADU and D-562 cells (0 ng, 100 ng and 200 ng), and changes in P62 protein expression were detected by western blot analysis. (H) Diagram of HOXC-AS2 gene body truncations: T1 and T2. (I) The T1 and T2 fragments of HOXC-AS2 were identified by 3% agarose gel electrophoresis. (J) The binding of HOXC-AS2 to the P62 protein was confirmed by a gene body truncation experiment. (K) The specific binding domain of HOXC-AS2 and P62 was confirmed by domain binding experiments.