Research Paper Volume 12, Issue 13 pp 12812—12840

Figure 6. The expression of PHLPP2 is regulated by LINC00402 and LINC00461 competitively binding to miR-141 as well as SFTA1P competitively binding to miR-424. (A) RT-qPCR was conducted to measure the expressions of lncRNAs when HT-29 cells were transfected with miR-141 inhibitor or mimic. (B) RT-qPCR was performed to measure the expressions of lncRNAs when the HT-29 cells were transfected with miR-424 inhibitor or mimic. (C) Dual-luciferase reporter gene assay verified LINC00402 and LINC00461 binding to miR-141. (D) Dual-luciferase reporter gene assay verified SFTA1P binding to miR-424. (E) FISH technology was utilized to identify the subcellular localization of LINC00402, LINC00461, and SFTA1P in the colon cancer (400 ×). (F) RNA pull-down was performed to show the relationship of LINC00402 and LINC00461 binding to miR-141, SFTA1P binding to miR-424. (G) LINC00402, LINC00461 and SFTA1P binding to Ago2 as reflected by the RIP assay. (H) Relative expression of PHLPP2 mRNA determined by RT-qPCR. (I) Protein expression of PHLPP2 protein normalized to GAPDH determined by Western blot analysis. * p < 0.05 vs. the NC group; # p < 0.05 vs. the PHLPP2 group; Measurement data in this Figure expressed as the mean ± standard deviation, while comparisons among multiple groups were conducted using One-Way ANOVA; the experiment was repeated three times independently.