Figure 6.

LINC00578 inhibits the ubiquitination of SLC7A11 by directly interacting with UBE2K. (a) Venn diagram showing 5 LINC00578 antisense proteins and 18 LINC00578 sense proteins by pull-down assay of PL45 cells. (b) Western blotting showing that LINC00578 interacts with UBE2K in pancreatic cancer cells. (c) qRT–PCR showing LINC00578 enrichment in UBE2K-immunoprecipitated RNAs in PL45 cells. An antibody against human IgG was used as a negative control for RIP. (d) qRT–PCR analysis showing the RNA level of SLC7A11 in the OE-LINC00578 group versus the vector group and the Sh-LINC00578 group versus the Sh-NC group. (e) Western blot assay revealing the reverse effect of MG132 on the protein degradation of SLC7A11 by Sh-LINC00578 introduction. (f) Western blot assay showing the promotion of CHX on the protein degradation rate of SLC7A11 by Sh-LINC00578 introduction. (g) Cells stably expressing Sh-NC or Sh-LINC00578 were subjected to a deubiquitination assay, and polyubiquitylated proteins were detected with an anti-Ub antibody. (h) Co-IP assay showing the interaction between UBE2K and SLC7A11. (i) IHC for SLC7A11 was applied between the LINC00578 high-expression group (above panel) and the low-expression group (below panel) in 50 paired pancreatic cancer tissue samples. The correlation analysis showed a positive correlation between LINC00578 and SLC7A11 (P = 0.0484, Fisher's exact test) expression. (j) Diagram illustrating how LINC00578 promotes SLC7A11 to inhibit ferroptosis. ^∗∗P < 0.01.