Fig. 3. Biochemical properties of a GFP-ZLAP2 fusion protein, and of full-length ZLAP2. A GFP-ZLAP2 fusion protein (amino acids 503-657 of ZLAP2; GFP503-657) expressed in Xenopus A6 cells (A), and ZLAP2 from zebrafish ovaries (B) was analyzed. The GFP was fused to the aminoterminus of this LAP2 deletion mutant. Proportional amounts of proteins of pellet fractions (P) and supernatants (S) were separated by SDS-PAGE and immunoblotted with GFP antibodies (A) or ZLAP2-specific antibodies (ZLAP2-serum1). The quality of the fractionation was controlled by immunoblotting with antibodies against lamin B2 (A', B'). Lamins should be recovered in the supernatant after urea extraction. (A, A') GFP-ZLAP2 fusion protein; Xenopus A6 cells extracted with 6 M and 8 M urea (A, GFP antibody; B', lamin B2 antibody). (B,B') Membranes of zebrafish ovaries purified by sucrose step gradient centrifugation were extracted with 4 M and 6 M urea. (B, ZLAP2-serum1; B', lamin B2 antibody). The positions of ZLAP2 (o) and ZLAP2 (g) are marked in (B) as well as molecular masses of reference proteins (in kDa).

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