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Fig. 2. Cytological characterization and effects of Cts7/Cts8
expression in TS cells. (A) Northern blot hybridization on
trophoblast stem (TS) cells grown in media promoting stem cell maintenance
(+FGF/CM) or differentiation (-FGF/CM). Expression of Cts7 and
Cts8 correlates with the profile of giant cell markers
(Cdx2, stem cells; Ascl2, ectoplacental cone and
spongiotrophoblast; Prl3d1, primary, parietal giant cells;
Prl3b1, secondary giant cells; Prl2c2, all giant cells).
(B) In situ hybridization on TS cell grown for 4 days in
differentiation medium. Some giant cells (blue, arrows) are positive.
(C) Immunostaining of CTS7 showing localization to the perinuclear
area, the Golgi, and to the cytoplasm in a granular pattern indicative of
endo- and lysosomal localization. (D) Western blots of transfected TS
cells and their supernatants. CTS7 and CTS8 are secreted into the medium;
intracellular control proteins (PFPL) were not detected in the supernatant.
(E,F) Relative cell size measurements of TS cells 2 days after
transfection with empty GFP-expression vector and Cts7-GFP (E) or
Cts8-GFP (F). Cathepsin expression causes a significant shift towards
larger cell sizes. Scale bars: 40 µm in B; 20 µm in C.
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