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Lineage-Restricted Expression of Protein Kinase C Isoforms in Hematopoiesis by Alessandra Bassini, Giorgio Zauli, Giovanni Migliaccio, Anna Rita Migliaccio,

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Presentation on theme: "Lineage-Restricted Expression of Protein Kinase C Isoforms in Hematopoiesis by Alessandra Bassini, Giorgio Zauli, Giovanni Migliaccio, Anna Rita Migliaccio,"— Presentation transcript:

1 Lineage-Restricted Expression of Protein Kinase C Isoforms in Hematopoiesis
by Alessandra Bassini, Giorgio Zauli, Giovanni Migliaccio, Anna Rita Migliaccio, Massimiliano Pascuccio, Sabina Pierpaoli, Lia Guidotti, Silvano Capitani, and Marco Vitale Blood Volume 93(4): February 15, 1999 ©1999 by American Society of Hematology

2 Western blot analysis of PKC-, βΙ , δ, ɛ, η, and ζ isoforms in homogenates obtained from freshly isolated CD34+ cells. Western blot analysis of PKC-, βΙ , δ, ɛ, η, and ζ isoforms in homogenates obtained from freshly isolated CD34+ cells. Isoform-specific antibodies were added to the membranes in the absence (left of each couple lanes) or presence of the corresponding immunizing peptide (right of each couple of lanes). A representative of three separate experiments is reported. Alessandra Bassini et al. Blood 1999;93: ©1999 by American Society of Hematology

3 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

4 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

5 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

6 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

7 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

8 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

9 Assay of PKC catalytic activity in homogenates obtained from various 32D subclones.
Assay of PKC catalytic activity in homogenates obtained from various 32D subclones. Data are reported as the means ± SD of three separate experiments performed in duplicate. Alessandra Bassini et al. Blood 1999;93: ©1999 by American Society of Hematology

10 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

11 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

12 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

13 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

14 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

15 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

16 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

17 Alessandra Bassini et al. Blood 1999;93:1178-1188
©1999 by American Society of Hematology

18 RT-PCR analysis of the expression of PKC-ɛ and - mRNA in the subclones of the 32D cell lines.
RT-PCR analysis of the expression of PKC-ɛ and - mRNA in the subclones of the 32D cell lines. The ethidium bromide staining of products amplified by 30 cycles of PCR with primers specific for the PKC-ɛ and - or for murine actin and separated by electrophoresis on agarose gel are presented. Comparable amounts of products of the expected molecular weights were amplified not only from cDNA obtained from human cord blood (last lane) and mouse bone marrow (first lane) mononuclear cells, as controls, but also from cDNA obtained from all subclones of the 32D cell lines. The specificity of the PKC-ɛ band amplified from the 32D Epo cell line was proved by direct sequencing of the band eluted from the gel. Alessandra Bassini et al. Blood 1999;93: ©1999 by American Society of Hematology


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