ZEB1-AS1 downregulates miR-200b , upregulates fascin-1 ( FSCN1 ), and promotes cell migration and invasion. a Luciferase assays were performed in T24, RT4, and 293 T cells co-transfected for 24 h with miR-negative control (NC) or miR-200b and a plasmid containing wild-type or mutant-type ZEB1-AS1 3′untranslated region (UTR) upstream the luciferase gene. Firefly luciferase activity of each sample was normalized by Renilla luciferase activity. Data were analyzed by T-test. b <t>RNA-binding</t> protein <t>immunoprecipitation</t> <t>(RIP)</t> assays with anti-AGO2 antibodies were performed in T24 and RT4 cells transiently transfected with miR-200b ; ZEB1-AS1 levels were detected by quantitative PCR (qPCR); 10% input was used as positive control and RIP with anti-IgG antibodies served as negative control. Glyceraldehyde 3-phosphate dehydrogenase ( GAPDH ) was used as the internal control. Data were analyzed by T-test. c Transfection efficiency of ZEB1-AS1 knockdown (si1, si2) and overexpression (OE) in T24 and RT4 cells was detected by qPCR. Data were analyzed by T-test. d and e The levels of miR-200b ( d ) and FSCN1 ( e ) were measured by qPCR after ZEB1-AS1 knocked down or overexpressed in T24 and RT4 cells. Data were analyzed by T-test. f E-cadherin, N-cadherin, vimentin, and FSCN1 protein expression in T24 and RT4 cells in which ZEB1-AS1 had been knocked down or overexpressed. Data were analyzed by T-test. ( g ) FSCN1 levels in T24 cells co-transfected with miR-NC or miR-200b and with an empty vector (EV) or a plasmid overexpressing ZEB1-AS1 . Data were analyzed by T-test. h and i Transwell assays (without or with Matrigel) to detect cell migration ( h ) and invasion ( i ) of T24 and RT4 after ZEB1-AS1 silencing or overexpression. j and k Transwell assays (without or with Matrigel) to detect cell migration ( j ) and invasion ( k ) of T24 and RT4 co-transfected with miR-NC or miR-200b and with an EV or a plasmid overexpressing ZEB1-AS1 . Data were analyzed by T-test. Data are presented as the mean ± standard deviation (SD). * P