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Fig. 5. Repression of c-Gcm1 targets inhibits neuronal differentiation without
affecting cell cycle exit. (A-O') Electroporation of E1.5
embryos with control (C,C',E,G,I-K,M,N) or c-Gcm1BD-ER (BD-ER;
A-B',D,D',F,H-J,L-O') expression vectors. Immunolabeling and
in situ hybridization on transverse sections were performed 24 hours after
electroporation using anti-ßIII-tubulin (Tuj1 in A,A') or Lim1/2
(B,B',E,F) antibodies or NeuroM probe (C-D').
(A-B') Overexpression of c-Gcm1BD-ER inhibits the generation of
terminally differentiated neurons. (E,F) High magnification of electroporated
neural tube showing that cells overexpressing c-Gcm1BD-ER do not express
Lim1/2, whereas a fraction of cells electroporated with a control vector
express Lim1/2 (arrow in E) as they reach the mantle layer. (I,J) Percentage
of transfected cells expressing Lim1/2 (I) or MNR2 (J) after electroporation
of control or c-Gcm1BD-ER expression vectors; asterisks indicate significant
difference (P<0.01). (G,H) BrdU (red) and GFP immunolabeling on
transverse sections obtained from embryos subjected to a 1 hour BrdU pulse
performed 24 hours after electroporation of control (G) or c-Gcm1BD-ER (H)
expression vectors. Note the presence of double-labeled cells in both cases
(arrows). (M,N) Percentage of transfected cells incorporating BrdU (M) or
expressing Sox3 (N) after electroporation of control or c-Gcm1BD-ER expression
vectors. Note that values are not significantly different. (K,L,O,O')
Similar profile of Sox3 (red in K,L) and Pax7 (red in O,O') expression
upon electroporation of control (K) or c-Gcm1BD-ER (L,O,O') expression
vectors. Scale bars: 50 µm in A-D',O,O'; 20 µm in
G,H; 60 µm in I,J; 40 µm in K,L.
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