13 In Situ Hybridization for RNA: Nonradioactive Probe: Oligo-DNA Probe
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standard protocol. A template DNA is necessary (Should be prepared by each researcher). Preparation of the tissue, hybridization and immunohistochemistry after hybridization are similar
to the methods described above. The protocol using double
strand DNA probe is described in --+ Chapter 9.
Results
Figure 1 shows the expression of mRNA for interleukin (lL)-4
and IL-4 receptor in glomeruli of a representative patient
with IgA nephropathy (Furusu et al. 1997). Note the clear staining of intraglomerular cells for IL-4 mRNA and IL-4 receptor
mRNA in perinuclear or cytoplasmic pattern. Examination of
mirror sections showed that some cells are double positive
for IL-4 mRNA and IL-4 receptor mRNA, while others are positive for each mRNA. As shown in the figure, the expression
Fig. 1. In situ detection ofIL-4 mRNA and IL-4 receptor mRNA in a renal tissue
sample from a representative patient with IgA nephropathy. Mirror sections of
glomeruli show that intraglomerular cells are clearly stained for IL-4 mRNA and
IL-4 receptor mRNA in perinuclear or cytoplasmic pattern. As shown here,
some cells are double positive for IL-4 mRNA and IL-4 receptor mRNA (arrowheads), while others are positivefor each mRNA (arrows). The expression of the
mRNAs is detected at a cellular level. Magnification x400.