Research Paper Volume 12, Issue 13 pp 12812—12840

PHLPP2 is regulated by competing endogenous RNA network in pathogenesis of colon cancer

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.