Project description:We have identified several nucleotide motifs (caug, cgggag=S2) that promote exosome sorting of miRNA in different cell types including brown adipocytes. In order to identify which proteins might recognize and bind to these motifs, we have performed co-precipitations of proteins binding biotinylated forms of miRNAs containing the aforementioned motifs or none - using streptavidin beads incubated with brown adipocytes cell lysates. We have included two types of controls: negative poly-A control and a scramble miRNA.
Project description:We have reported that microRNAs are present in human, bovine, and rat milk whey. Milk whey miRNAs were resistant to acidic condition and to RNase. Thus, milk miRNAs were thought to be present packaged into membrane vesicles like exosome. However, body fluid miRNAs have been reported that there are in different forms. To clarify which miRNAs species are exist in exosome and which species are exist in another form, we used bovine raw milk and purified total RNA from exosome fraction and ultracentrifugated supernatant fraction, and analyzed by miRNA microarray.
Project description:We explored the biological effects of AGO2 downregulation on the expression of miRNAs in human NSCLC cells. Using AGO2 siRNA as a positive control, we overexpressed miR-138 in H1299 cells to observe changes in the miRNA profile using an miRNA microarray assay.
Project description:We explored the biological effects of AGO2 downregulation on the expression of miRNAs in human NSCLC cells. Using AGO2 siRNA as a positive control, we overexpressed miR-138 in H1299 cells to observe changes in the miRNA profile using an miRNA microarray assay. Total RNAs were extracted from H1299 cells transfected with NC, AGO2 siRNA or miR-138 for 48h. Their microRNAs expression profiles were investigated using LCSciences Human miRNA Array platform (LCSciences, Houston, TX; Sanger mirBase Release 19.0).
Project description:We used the NanoString Human nCounter miRNA expression platform to compare the miRNA expression profiles of QKI-5 knockdown H1299 cells and control cells We found that QKI-5 knockdown significantly changed miRNA expression pattern in lung cancer cells
Project description:MiR-138 has a variety of biological functions because of its capacity to act on different target genes in various cells and tissues; however, the targets of miR-138 in human non-small cell lung cancer cell line H1299 cannot be determined by bioinformatics alone. Thus, H1299 cells overexpressing miR-138 in H1299 cells were subjected to microarray analysis to analyse the differences of gene expression.