Fig. 3

LINC00689 sponges with miR-496 in prostate cancer. a The binding sites of LINC00689-WT, miR-496 and LINC00689-Mut predicted by starbase were displayed. b Luciferase reporter assay examined the luciferase activity of LINC00689-WT/Mut in miR-496 mimics transfected DU145 and LNCaP cells. c RT-qPCR measured the expression of miR-496 in sh-LINC00689#1 transfected DU145 and LNCaP cells (post 48 h of transfection). d The expression of miR-496 in normal prostate epithelial cell (RWPE1) and PCa cells (DU145, LNCaP, PC-3 and C42B) was detected by RT-qPCR. e The knockdown efficiency of miR-496 was measured in miR-496 inhibitor transfected DU145 and LNCaP cells (48 h for plasmid transfection). f, g MTT (for indicated time) and colony formation (14 days later) assays detected DU145 cell proliferation in differently transfected groups (sh-NC, sh-LINC00689#1 and sh-LINC00689#1 + miR-496 inhibitor). h Flow cytometry analysis examined DU145 cell apoptosis in differently transfected groups (sh-NC, sh-LINC00689#1 and sh-LINC00689#1 + miR-496 inhibitor). i Western blot assay measured cleaved caspase 3 expression in differently transfected groups (sh-NC, sh-LINC00689#1 and sh-LINC00689#1 + miR-496 inhibitor). GAPDH was an internal control. j Caspase-3 test measured DU145 cell apoptosis in differently transfected groups (sh-NC, sh-LINC00689#1 and sh-LINC00689#1 + miR-496 inhibitor). k, l Transwell assay (24 h) measured DU145 cell migration and invasion in differently transfected groups (sh-NC, sh-LINC00689#1 and sh-LINC00689#1 + miR-496 inhibitor). Images were captured by using the inverted microscope (4 × objective lens) (scale bar = 100 μm). Error bars represent the mean ± SD of at least three independent experiments. ^**P < 0.01