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. 2008 Jan;19(1):262–273. doi: 10.1091/mbc.E07-08-0798

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© 2007 by The American Society for Cell Biology

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Figure 5. — Immunoblot analysis of HA-tagged Uni2 protein and phosphatase-treated Uni2 protein. Proteins were separated by SDS-PAGE and transferred to a PVDF membrane. A high-affinity anti-HA antibody was used to identify the HA-tagged Uni2 protein. (A) Protein extracts were lane 1, WT; 2, uni2-3; and 3–4, two independent uni2-3 strains phenotypically rescued with the HA-tagged UNI2 gene. Top, a high-affinity anti-HA antibody was used to identify the HA-tagged Uni2 protein; bottom, an antibody against β-tubulin was used as a loading control. (B) Protein from an HA-tagged transformant after a 30-min treatment at 37°C with CIP buffer and the indicated reagents. The blot was probed with the anti-HA antibody.