Regulation of chicken IL-2 expression by miR-26a in DF-1 cells. (a) Northern blot analysis of total RNA extracted from DF-1 cells transfected with miR-26a expression or control vectors using anti-miR-26a probe. (b) Northern blot analysis demonstrating the specific silencing of miR-26a expression with anti-miR-26a, but not with anti-miRNA control. U6 RNA was used as the control probe for loading control. (c) Western blot analysis of chicken IL-2 expression in DF-1 cells co-transfected with HA-tagged IL-2 expression construct pcDNA-IL-2-3HA-3'UTR and miR-26a expression vector (lane 2) or control vector (lane 3) using anti-HA tag antibody. (d) Western blot analysis of the IL-2 levels in the presence of anti-miR-26a or anti-miRNA control with anti-HA tag antibody. The levels of α-tubulin in the cell lysates are also shown as the loading control. Quantitation of the band densities was carried out using ImageQuant 300 software.