Journal: bioRxiv

Article Title: Maize RAMOSA3 accumulates in nuclear condensates enriched in RNA POLYMERASE II isoforms during the establishment of axillary meristem determinacy

doi: 10.1101/2021.04.06.438639

Figure Lengend Snippet: A) Representative images of whole-mount immunolocalization in maize inflorescences showing the expected pattern for RA3 (green specles) subtending the base of SPMs. B) ra3 mutant was used as a negative control, showing the absence of signal C) RA3 (green) is localized in nuclear and cytoplasmic speckles, the nuclear envelope was labeled using an anti-Nuclear Pore Complex protein antibody (anti-NPC, in red), the nucleus shape is highlighted by the white dashed line, a representative median confocal section shows the nucleus surrounded by the NPC, with RA3 nuclear speckles inside. Sections were counterstained with DAPI (blue). D) Axillary spikelet pair meristem (SPM) development in ears occurs continuously on the flanks of the Inflorescence Meristem (IM), image on left is a scanning electron micrograph; whole-mount double immunolocalizations were performed using 2-5 mm ears. 3D Z-stacks of nuclei were acquired from the uppermost three SPMs (marked orange on the SEM) in cells expressing RA3 and covering entire nuclei (~5 μm). E) after acquisition, images were processed by deconvolution and cropping a 3D region of interest inside the nucleus (dashed lines) and the percentage colocalization was calculated by volume (object based) and rendered for easy representation and visualization. Scale bars, A-C and E : 2 μm, D : 20 μm.

Article Snippet: Additional images, including anti-NPC, were acquired using a Zeiss LSM 900 using the Airyscan 2 mode under equivalent parameters, or using 3D Structured Illumination Microscopy on a DeltaVision OMX super-resolution microscope under a 100X objective.

Techniques: Mutagenesis, Negative Control, Labeling, Expressing