Subcellular localization of PRN1 in dark-grown seedlings.

Constructs and transgenic plants are described in methods. 6-d-old dark-grown prn1 whole seedlings transformed with PRN1::PRN1-GFP (T3) or WT seedlings transformed with 35S::PRN1-GFP were fixed, stained with DAPI, mounted on slides, then photographed on a spinning disk confocal using steady state lasers 405 nm, 488 nm, and 561 nm. All images shown are from the cotyledon epidermis layer. Panel rows are indicated by wavelength, with individual channels in black and white. In the merge, 405 nm (DAPI) is false-colored blue, 488 nm (GFP) is false-colored green, and 561 nm is false-colored red (Red). Images are representative with no alteration of the fluorescence within the image field, and images represent an optical section of 1 μm thickness. The column heading indicates the different seedling lineages. Yellow boxes on the figure indicate areas of interest and show an enlargement. Red arrows indicate DAPI stain in nucleus. White arrows indicate GFP accumulation in 488 nm. Untransformed WT and prn1 seedlings are also shown on the figure. Scale bars each represent 25 μm. n = 4 biological replicates, with at least 12–20 individual seedlings viewed per replicate; images are representative.