| |
Fig. 3. (A) Total cellular proteins of HeLa cells (HeLa, lane 1),
Xenopus cultured A6 cells (A6, lane 2), and Xenopus tissues (lane 3,
follicle cells [Fol]; lane 4, spleen; lane 5, liver) after separation by
SDS-PAGE (12% acrylamide) and immunoblotting with affinity
purified antibodies of the MAN-serum specific for LAP2/TP
proteins. Antibodies had been affinity purified on the 187 aminoterminal
amino acids of the rat LAP2b. (B) Comparison of the
electrophoretic mobility of in vitro translated XLAP2 (lane 1) with
the major polypeptide detected by immunoblotting with the whole
MAN-serum in Xenopus follicle cells (lane 2). The in vitro
translation products were synthesized by coupled in vitro
transcription and translation of the cDNA coding for XLAP2.
Polypeptides of the two samples shown in B were separated in
neighbouring lanes in the same SDS-PAGE, and transferred to
nitrocellulose. The nitrocellulose strip containing lane 1 was directly
processed for fluorography, whereas the second strip (lane 2) was
incubated with the whole MAN-serum. Molecular masses of
reference proteins (in kDa) are marked in A and B. (C,C¢) Indirect
immunofluorescence microscopy (C) on Xenopus A6 cells with
affinity purified LAP2/TP antibodies of the MAN-serum. Antibodies
had been affinity purified on the full-length rat LAP2. (C¢) Staining
with the DNA dye Hoechst. Bar, 10 mm. |
