Figure 11. Effect of MALAT1 on PNPO and miR-216b-5p expression in breast cancer cells. (A) Identification of the sequence of binding sites between MALAT1 and miR-216b-5p. MALAT1-wt, wild-type MALAT1; MALAT1-mut, mutated MALAT1. (B) Detection of luciferase activity by the dual-luciferase reporter assay in HEK-293T cells. (C) Detection of MALAT1 expression by qRT-PCR in MCF-7 cells after si-MALAT1 treatment. (D) Detection of miR-216b-5p expression by qRT-PCR in MCF-7 cells after si-MALAT1 treatment. (E) Detection of MALAT1 expression by qRT-PCR in MDA-MB-231 cells after si-MALAT1 treatment. (F) Detection of miR-216b-5p expression by qRT-PCR in MDA-MB-231 cells after si-MALAT1 treatment. (G) Effect of miR-216b-5p on MALAT1-mediated PNPO expression. HEK-293T cells were treated with si-NC plus miR-inhibitor-NC, si-MALAT1 along, miR-216b-5p along, or si-MALAT1 plus miR-216b-5p for 48 h. The expression of PNPO protein was detected by Western blot (upper panel) and semi-quantification (lower panel). miR-NC, negative control of miRNA; miR-inhibitor-NC, negative control of miR-inhibitor; si-NC, negative control of siRNA; si-MALAT1, MALAT1-siRNA. n = 3; * P < 0.05.