Fig. 7
MALAT-1 and miR-200c-3p are associated with the immunoprecipitated AGO2 complex. a Western blot was employed to detect AGO2 expression after AGO2 knockdown in AsPC-1 and CFPAC-1 cells. b, c RT-qPCR was use to examine MALAT-1 (b) and miR-200c-3p (c) level in response to knockdown of AGO2 in AsPC-1 and CFPAC-1, respectively. d Binding sites for miR-200c-3p in MALAT-1, as predicted by Starbase v2.0. The parts in the column imply the possible binding sites in MALAT-1. e Seed matches from MALAT-1 wt or its mutants (MALAT-1-mut-1 and MALAT-1-mut-2) that were devoid of specific miR-200c-3p-binding sites were mutagenized and transfected into 293Â T cells together with 50Â nM of miR-200c-3p mimics or nc. MiR-200-3p mimics significantly decreased luciferase activity only in the MALAT-1-wt construct. f, g RIP with monoclonal anti-Ago2 and IgG from AsPC-1 (f) and CFPAC-1 (g) cell extracts transfected with nc or miR-200c-3p mimics. Levels of MALAT-1 and miR-200c-3p were measured by RT-qPCR