|
|
|
|
|
|
|
||||
| Home Help Feedback Subscriptions Archive Search Table of Contents | ||||
|
Fig. 7. Reduction of cell proliferation and neurogenesis by persistent
Hes1 expression. (A) The schematic structures of
recombinant retroviruses. (B-G) Immunostaining for GFP. E11.5
telencephalic neural progenitors infected with CLIG or CLIG-Hes1 were examined
at different time points. (H) Clonal sizes (with s.e.m.) of the
virus-infected cells. Clonal sizes of CLIG-Hes1-infected cells are smaller
than those of CLIG-infected cells. Experiments were repeated at least six
times at each time point. (I) Ratios (with s.e.m.) of TUNEL-positive
cells in the virus-infected cells at 72 hours after infection (at least 2,000
cells in three independent experiments were examined). Although there is a
tendency for cell death to increase in CLIG-Hes1 infection compared to CLIG
infection, the effect is not sufficient for the reduction of growth rates in
CLIG-Hes1 infection. (J) Ratios (with s.e.m.) of proliferating cells in
the virus-infected cells at 72 hours after infection (at least 500 cells in
three independent experiments were examined). Ki67-positive cell ratios are
similar between CLIG- and CLIG-Hes1-infected cells, whereas a ratio of cells
positive for the G1 phase-specific cyclin D1 is higher in CLIG-Hes1 infection,
suggesting that Hes1 overexpression prolongs G1 phase. (K-Z)
Immunocytochemical staining of cells infected with CLIG (K-N,S-V) and
CLIG-Hes1 (O-R,W-Z). After 5 days in culture, some CLIG-infected cells have
differentiated into TuJ1-positive neurons (K-N) or GFAP-positive astrocytes
(S-V) whereas virtually none of the CLIG-Hes1-infected cells (closed
arrowheads) are neurons (O-R) or astrocytes (W-Z). Some neurons (TuJ1-positive
cells) and astrocytes (GFAP-positive cells) are indicated by open arrowheads
(R,V).
|
