5 Introduction to the Detection of Specific DNA and RNA Sequences
59
G
e
Fig. 2. An example of a set of control experiments (Koji and Brenner 1993). The
set was performed to verify the specificity of the signals for estrogen receptor
(ER) mRNA in the frozen sections of monkey uterus. (a) Antisense: A section
was hybridized with Dig-ER antisense oligo-DNA. Strong staining for ER mRNA
was found in glandular epithelial cells as well as stromal cells. (b) Sense: An
adjacent section to the section hybridized with the antisense probe was hybridized with Dig-ER sense oligo- DNA. No staining was found. (c) Competition:
An adjacent section was hybridized with Dig-ER antisense probe in the presence
of an excess amount (33-fold) of non-labeled ER antisense oligo-DNA. The signal for ER mRNA was markedly reduced. (d) Non-competition: When an adjacent section was hybridized with Dig-ER antisense probe in the presence of an
excess amount (33-fold) of non-labeled unrelated oligo-DNA. The intensity of
ER mRNA signal was not changed, compared to that of "competition test". (e)
RNase: A section was digested with RNase A before hybridization, and then
hybridized with Dig-ER antisense probe. The section was not stained essentially.
G; gland, S; stroma