Fig. 2. Localization of the Y23K97 chimeric protein by indirect immunofluorescence. The staining of SNY9-1 [
pep4 (A,B,C,D,E,F)] and SNH6-1C-3.1 cells [rer1/pep4 (G,H,I,J,K,L,M,N)] overexpressing Mns1p (A,B,G,H,I), Y23K97 (C,D,J,K,L) or Kre2p (E,F,M,N) was performed with affinity-purified anti-Mns1p antibodies (A,G) or affinity-purified anti-Kre2p antibodies (C,E,J,M) followed by TRITC-conjugated secondary antibodies. In rer1/pep4 cells, double labeling with anti-Mns1p and anti-vacuolar H+-ATPase antibodies (G,H) or anti-Kre2p and anti-H+-ATPase antibodies (J,K) was performed in the same cells. Endogenous vacuolar H+-ATPase was detected using a monoclonal antibody followed with CY2-conjugated secondary antibodies (H,K). DAPI staining of the nuclei is shown (B,D,F,I,L,N). Parallel arrows in figures G,H,I and in figures E and F indicate double-labeling of the same cells. (Bar, 5 µm).
