CG&D
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cell Growth & Differentiation

Right arrow Help viewing high resolution images
Right arrow Return to article


Fig. 3. Expression and characterization of Mi proteins. A, characterization of the serum directed against Mi protein. The murine mi cDNA inserted in the pSG5 expression vector was translated in [35S]methionine-containing reticulocyte lysate. Translated proteins were immunoprecipitated with rabbit serum MiC prepared against bacterially expressed Mi COOH-terminal peptide (Lane 1) and with cognate preimmune serum (Lane 2). Lane 3, translated crude Mi protein. B, detection of Mi by immunoprecipitation. Selected LE-mi-transfected CNR cells or LE-transfected CNR cells were labeled with [35S]methionine/cysteine for 30 min. Labeled cells were solubilized in RIPA buffer. Cell lysates were then immunoprecipitated with the serum MiC (Lanes 2 and 4) and with cognate preimmune serum (Lanes 3 and 5). Lane 1, LE-mi-CNR lysate were first immunoprecipitated with the serum MiC, and proteins recovered from the immunocomplexes bound to protein A-Sepharose were immunoprecipitated again with the same serum. C: a, phase contrast of selected LE-mi-CNR. b, subcellular localization of Mi assayed by indirect immunofluorescence on fixed cells with serum MiC. Anti-Mi-immunoreactive proteins were detected with tetramethylrhodamine isomer R-labeled swine antirabbit immunoglobulins secondary reagent. c, phase contrast of selected LE-CNR used as control. d, subcellular localization of Mi assayed by indirect immunofluorescence. Nuclei are labeled in LE-mi-CNR cells but not in control cells.





Right arrow Return to article


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cell Growth & Differentiation