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Fig. 2. p38 and JNK are active during mouse preimplantation development. (A) The
activated form of p38 was detected by using anti-phospho-specific p38
MAP kinase antibody. Embryos at the four-cell, eight-cell, morula or
blastocyst stage were stained with anti-phospho-specific p38 MAP kinase
antibody. Fluorescence was viewed with a confocal microscope. (B)
Phosphorylation of MAPKAPK-2 or HSP27 with or without SB203580. The eight-cell
stage embryos were treated with SB203580 for 1.5 hours, and then the embryos
were fixed and stained with anti-phospho-specific MAPKAPK-2 (Thr 334) antibody
or anti-phospho-specific HSP27 (Ser 82) antibody. (C) Activation of JNK was
assessed by using anti-phospho-specific Jun (Ser 73) antibody. Embryos at the
four-cell, eight-cell, morula and blastocyst stage were stained with
anti-phospho-specific Jun (Ser 73) antibody. Fluorescence was viewed with a
confocal microscope. (D) Phosphorylation of Jun was examined with or without
SP600125. SP600125 was added at the eight-cell stage. After 2 hours of
incubation, the embryo was fixed and stained with anti-phospho-specific Jun
(Ser 73) antibody. (E) Effect of JNK1/2-targeting siRNAs on mouse
preimplantation development. One-cell stage embryos were not injected or
injected with DDW or dsRNA oligonucleotides as indicated, and cultured for 3
days. The numbers of morphologically normal or abnormal embryos were counted.
Bright-field microscopic photographs are shown.
