Figure 4. Co-immunoprecipitation of pre-mature miR-200s with XPO₅ in tumoral pancreatic cell lines.

(A) nuclear (N) and cytoplasmic (C) fractions prepared from tumoral pancreatic cell lines were separated by electrophoresis on acrylamide gel and XPO₅ detected by immunoblotting. Cytoplasmic and nuclear preparations were verified by immunoblotting of calnexin and histone H1, respectively. Data are representative of 2 separate experiments. (B) XPO₅ was immunoprecipitated in cytoplasmic and nuclear fractions prepared from Mia-Paca2 and BxPC3 cells. Premature miRNAs that co-immunoprecipitate with XPO₅ were extracted then measured by RT-qPCR. In parallel, total pre-mature miRNAs level present in cytoplasmic and nuclear fractions was determined prior to immunoprecipitation. As control, immunoprecipitation with irrelevant IgG was performed. Each value is the mean ± SD of triplicates. Data are representative of 2 separate experiments. Data are expressed as the ratio between immunoprecipitated and total pre-mature miRNA determined for each cell fraction. pre-miR-141, pre-mature miR141; pre-miR-429, pre-mature miR429.