Mouse anti α acetylated tubulin antibody

The following primary antibodies were used: mouse anti-α-acetylated-tubulin antibody (Santa Cruz Biotechnology; 1:1000), mouse anti-Islet-1 antibody (DSHB; 1:200), mouse anti-Zn-8 antibody (ZIRC; 1:200), rabbit anti-glial brillary acidic protein (Sigma; 1:500), mouse anti-Zpr-1 antibody (ZIRC; 1:200), mouse anti-HA antibody (Santa Cruz Biotechnology; 1:100), and mouse anti-GFP antibody (Molecular Probes; 1:200).
Cell culture, transfection. and immuno uorescence RPE1 cells were cultured as described previously [30] . For transient expression, RPE1 cells were transfected with Lipofectamine 2000 (Invitrogen) according to the manufacturer's protocol. At 24 h posttransfection, cells were processed as described for each experiment. Cells were xed in 4% paraformaldehyde in phosphate-buffered saline (comprising 20 mM sodium dihydrogen phosphate, 0.9% sodium chloride [NaCl], pH 7.4), permeabilized with 0.3% Triton X-100, and labeled with primary antibodies for 1 h at room temperature followed by incubation with appropriate secondary antibodies conjugated to Alexa 488 or Alexa 568.