Fig. 2

Overexpression of miR-214-3p promotes porcine GC proliferation. MiR-214-3p agomir or negative control (NC) transfected into cells harvested after 24 h. a Overexpression efficiency of miR-214-3p after transfection with miR-214-3p agomir compared to NC; b Flow cytometry determines cell percentages in different cycle phases; c Cell cycle analysis statistical results; d EdU staining assay of proliferous cell quantities. Positive cells stained by EdU (red) and total cell nucleus stained with Hoechst (blue); e Results presented as red/blue cell nuclei; f CCK-8 assay detects cell viability after 24-h transfection as absorbance value at 450 nm after incubation with 10% CCK-8 solution for 4 h; g RT-qPCR detects cell cycle genes, Cyclin B, Cyclin E, Cyclin D, CDK4 after 24-h transfection; h Western blot analysis of cell cycle genes; i Quantification of Western blot analysis of Cyclin B, Cyclin D, Cyclin E, CDK4. Note: Data are mean ± SEM of three independent experiments; * P < 0.05, ** P < 0.01