Fig. 4.
Fig. 4. MEFV expression in peripheral blood leukocytes. / (A) EtBr-stained PAGE of RT-PCR products of the MEFV and β-actin mRNAs from highly purified populations of peripheral blood neutrophils (lane 1), eosinophils (lane 2), macrophages and monocytes (lane 3), and lymphocytes (lane 4). Negative and positive control PCR reactions using water and the MEFV-containing plasmid pV75-1, respectively, are shown (lanes 5 and 6). (B) Purity of populations analyzed in A, as determined by fluorescence-activated cell sorting. Cells were stained for a cell-type–specific marker (red) and nonspecific isotype control antibody (green). The antibodies used were monocytes (anti-CD14), neutrophils (anti-CD16), B cells (anti-CD20), and T cells (anti-CD3). (C) Wright-Giemsa–stained cytospin preparations of eosinophils used for these analysis. Original magnification ×400. (D) Photomicrographs of Wright-Giemsa–stained monocyte in situ hybridizations. Results with use of a gene-specificMEFV antisense riboprobe (right panel) and a nonspecificMEFV sense strand control riboprobe (left panel) are shown. Original magnification ×400.

MEFV expression in peripheral blood leukocytes.

(A) EtBr-stained PAGE of RT-PCR products of the MEFV and β-actin mRNAs from highly purified populations of peripheral blood neutrophils (lane 1), eosinophils (lane 2), macrophages and monocytes (lane 3), and lymphocytes (lane 4). Negative and positive control PCR reactions using water and the MEFV-containing plasmid pV75-1, respectively, are shown (lanes 5 and 6). (B) Purity of populations analyzed in A, as determined by fluorescence-activated cell sorting. Cells were stained for a cell-type–specific marker (red) and nonspecific isotype control antibody (green). The antibodies used were monocytes (anti-CD14), neutrophils (anti-CD16), B cells (anti-CD20), and T cells (anti-CD3). (C) Wright-Giemsa–stained cytospin preparations of eosinophils used for these analysis. Original magnification ×400. (D) Photomicrographs of Wright-Giemsa–stained monocyte in situ hybridizations. Results with use of a gene-specificMEFV antisense riboprobe (right panel) and a nonspecificMEFV sense strand control riboprobe (left panel) are shown. Original magnification ×400.

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