Supplementary Figure 1: Expected results for variations in the tissue permeabilization and probe hydrolysis steps.
From: Analysis of 3D gene expression patterns in plants using whole-mount RNA in situ hybridization
(a–b) Examples of whole-mount in situ experiments to localize WUSCHEL mRNA in tissue that has been subjected to enzymatic permeabilization of either 0 min (a) or 6 min (b). Compare the absent or reduced signal in the thicker parts of the tissue, such as the center of the SAM (red arrowheads) or in older flowers, either at stage 3 (blue arrows) or stage 2 (red arrows). (c) Lane 1 shows the products of a transcription reaction (for 1 μl of the 20 μl reaction volume) from a PCR-amplified full-length (291 bp) CLAVATA3 template. Note the presence of an additional larger band (solid triangle), which is visible in most transcription reactions. Lane 2 shows the results of a successful hydrolysis reaction (for 5 μl of the 80-100 μl reaction volume), where a smear centered at roughly 0.1 kb is typically observed (open triangle). In the case of an unsuccessful hydrolysis, lane 2 would look identical to lane 1.
