Fig. 4

Mir100hg augmented lung cancer metastasis by targeting miR-15a-5p/miR-31-5p. A Mir100hg was localized in both the nucleus and cytoplasm as determined by FISH (Mir100hg, Red); B The localization of Mir100hg in both nucleus and cytoplasm was confirmed by RNA separation assay(Actin was used as a cytoplasmic control and U6 as a nuclear control; C Direct binding of Mir100hg to AGO2 was determined by RIP; D Schematic representation of Mir100hg targeting miR-15a-5p and miR-31-5p inferred from Mir100hg sequence and sequencing data from LLC-SD and LLC; E 37 candidate miRNAs screened from Annolc and sequencing data F The expression of miRNAs was examined by RT-PCR in OE-Mir100hg-LLC and sh-Mir100hg-LLC cells; G Luciferase assay confirmed that Mir100hg could targeted miR-15a-5p and miR-31-5p directly; H The efficiency of overexpression of miR-15a-5p and miR-31-5p and co-overexpression of the two miRNAs in OE-Mir100hg-LLC cells; I-J Migration and invasion of co-overexpression of miR-15a-5p and miR-31-5p in OE-Mir100hg-LLC cells; K Lung metastasis of mice was observed after tail vein injection of OE-miR-15a-5p + 31-5p-OE-Mir100hg-LLC (red); L-M HE staining was used to observe and quantify lung metastases. (*p < 0.05, **p < 0.01, ***p < 0.001)