Specificity testing of AZIN2 antisera.

<p>(A) Immunoblot from COS-7 cells transfected with plasmids carrying FLAG-tagged AZIN1, AZIN2, or ODC gene or an empty FLAG-vector (Ctrl). An Odyssey Infrared Imager was used for simultaneous two-color detection of rabbit anti-AZIN2 (antiserum 3) and mouse anti-FLAG antibodies. The lowest panel represents merged blots, yellow indicating co-localization of anti-AZIN2 and anti-FLAG. (B) Confocal laser scanning microscopy of COS-7 cells expressing one of the following genes: AZIN2-FLAG, AZIN1-FLAG, or ODC-FLAG. The cells were stained with either AZIN2 antiserum 2 or 3 together with mouse anti-FLAG, followed by donkey anti-rabbit Alexa Fluor-555 and donkey anti-mouse Alexa Fluor-488. AZIN2-FLAG-transfected cells were also stained with preimmune sera of the corresponding rabbits. In the merged images, yellow indicates the co-localization. Bars represent 10 µm.