Biotin signals were visualized using avidin-DCS rhodamine (A2012, Vector Labs), followed by biotinylated anti-avidin antibody (Ab73235, Abcam) plus an additional incubation with the avidin-DCS rhodamine, each used at 1:100 dilution in 1% BSA, 4 Ă—SSC, 0.2% Tween-20.