Phosphorylation at Serines 216 and 221 Is Important for Drosophila HeT-A Gag Protein Stability
Figure 3
Confirmation of stable cell lines expressing HeT-A Gag proteins.
(A) The stable cell lines were confirmed by isolating genomic DNA from cells and using a vector specific primer (PMK33-1) and an insert-specific primer (9D4 R4) by PCR. 10 ul of PCR product was run on 1% agarose gel and stained with ethidium bromide and photographed (Lane 1 = Pos, Lane 2 = S2 cells only, Lane 3 = Vector alone, Lane 4 = wild type HeT-A Gag, Lane 5 = M1 Gag, Lane 6 = M2 Gag, Lane 7 = M3 Gag). (B) Wild type and mutant HeT-A Gag stable cell lines expressed the same size Gag proteins as confirmed by immunoblot. Expression was only seen when cells were induced with CuSO4 (lane 1 = S2 cell, Lane 2 = Vector alone, Lane 3 = wild type Gag, Lane 4 = M1 Gag, Lane 5 = M2 Gag, Lane 6 = M3 Gag. (C) In wild type and mutant stable cell lines Gag protein was localized in the nuclear fraction as confirmed by immunoblot.