On page 850 in the February 1, 2000, issue, there are errors in Figure 5. The authors have provided a replacement Figure 5, incorporating a new image for the Western blot for Cdk2 after immunoprecipitation of Cdk2. The original Figure 5 IP Cdk2/WB Cdk2 used in the Blood manuscript was inadvertently used in a figure representing a different experiment in another manuscript. During the investigation of this problem, a validated Cdk2-IP/WB gel was located and has now been used to replace the row in question. In addition, black lines have been added to the row showing the IP + WB for cyclin E and the row showing the kinase assay, per Blood's current policies, to indicate that blank lanes were spliced out. The authors removed the quantification under the lanes and revised the figure legend by deleting the last sentence, which read, “Relative protein levels/kinase activity values are shown below all lanes (PLL-nonadherent is arbitrarily 1).” The correct figure and legend are shown.
![Figure 5. Adhesion to FN. Adhesion to FN is associated with increased levels of p27KIP1 and decreased cdk2-kinase activity. Mobilized blood CD34+ cells cultured for 48 hours in serum-free medium with low-dose cytokines were incubated with 8A2, washed, and plated in PLL- or FN-coated dishes for 12 to 16 hours. Adherent (Adh) and nonadherent (NA) cells were collected separately, and cells were lysed. A representative example of 3 individual experiments is shown. For p27KIP1, protein extracts were separated by SDS-PAGE, transferred onto nitrocellulose, and incubated with antibodies against p27KIP1 and goat anti–mouse HRP-conjugated antibody. Cyclin-E was immunoprecipitated from protein G–agarose beads. Immune complexes were separated by SDS-PAGE and blots probed with anti–cyclin-E antibodies and goat anti–mouse HRP-conjugated antibody. Protein bands were visualized with the use of the ECL detection system, and cdk2 was immunoprecipitated with the use of protein G–agarose beads. The immune complexes were separated by SDS-PAGE, and blots were probed with anti-cdk2 antibodies and goat anti–mouse HRP-conjugated antibody. Cdk2 activity was assayed by adding 5 μg histone and 10 μCi [r-32P]. Reaction products were resolved by SDS-PAGE, and the gel was exposed to X-ray film. Differences in protein levels were evaluated by scanning images with a GS-700 Imaging Densitometer and quantitated with the use of Molecular Analyst software.](/view-large/figure/8163078/zh89990939850001.jpeg)
Adhesion to FN. Adhesion to FN is associated with increased levels of p27KIP1 and decreased cdk2-kinase activity. Mobilized blood CD34+ cells cultured for 48 hours in serum-free medium with low-dose cytokines were incubated with 8A2, washed, and plated in PLL- or FN-coated dishes for 12 to 16 hours. Adherent (Adh) and nonadherent (NA) cells were collected separately, and cells were lysed. A representative example of 3 individual experiments is shown. For p27KIP1, protein extracts were separated by SDS-PAGE, transferred onto nitrocellulose, and incubated with antibodies against p27KIP1 and goat anti–mouse HRP-conjugated antibody. Cyclin-E was immunoprecipitated from protein G–agarose beads. Immune complexes were separated by SDS-PAGE and blots probed with anti–cyclin-E antibodies and goat anti–mouse HRP-conjugated antibody. Protein bands were visualized with the use of the ECL detection system, and cdk2 was immunoprecipitated with the use of protein G–agarose beads. The immune complexes were separated by SDS-PAGE, and blots were probed with anti-cdk2 antibodies and goat anti–mouse HRP-conjugated antibody. Cdk2 activity was assayed by adding 5 μg histone and 10 μCi [r-32P]. Reaction products were resolved by SDS-PAGE, and the gel was exposed to X-ray film. Differences in protein levels were evaluated by scanning images with a GS-700 Imaging Densitometer and quantitated with the use of Molecular Analyst software.
Adhesion to FN. Adhesion to FN is associated with increased levels of p27KIP1 and decreased cdk2-kinase activity. Mobilized blood CD34+ cells cultured for 48 hours in serum-free medium with low-dose cytokines were incubated with 8A2, washed, and plated in PLL- or FN-coated dishes for 12 to 16 hours. Adherent (Adh) and nonadherent (NA) cells were collected separately, and cells were lysed. A representative example of 3 individual experiments is shown. For p27KIP1, protein extracts were separated by SDS-PAGE, transferred onto nitrocellulose, and incubated with antibodies against p27KIP1 and goat anti–mouse HRP-conjugated antibody. Cyclin-E was immunoprecipitated from protein G–agarose beads. Immune complexes were separated by SDS-PAGE and blots probed with anti–cyclin-E antibodies and goat anti–mouse HRP-conjugated antibody. Protein bands were visualized with the use of the ECL detection system, and cdk2 was immunoprecipitated with the use of protein G–agarose beads. The immune complexes were separated by SDS-PAGE, and blots were probed with anti-cdk2 antibodies and goat anti–mouse HRP-conjugated antibody. Cdk2 activity was assayed by adding 5 μg histone and 10 μCi [r-32P]. Reaction products were resolved by SDS-PAGE, and the gel was exposed to X-ray film. Differences in protein levels were evaluated by scanning images with a GS-700 Imaging Densitometer and quantitated with the use of Molecular Analyst software.
This feature is available to Subscribers Only
Sign In or Create an Account Close Modal