Local chromatin environment of a Polycomb target gene instructs its own epigenetic inheritance
- John Innes Centre, United Kingdom
Figures
Figure 1 with 1 supplement
FLC-Venus and FLC-mCherry plants are late flowering and vernalization responsive.
(A) Schematic of FLC genomic DNA used to generate FLC-Venus and FLC-mCherry translational fusions. Exons are represented by black boxes. Transgenes extend from 3.4 kb upstream to 8.6 kb downstream …
Figure 1—figure supplement 1
Detection of FLC-Venus protein.
(A) Immunoblot of FLC-Venus GFP-trap immunoprecipitation (IP). wt indicates untransformed wild-type control sample (Col-FRI), whereas Venus indicates FLC-Venus transgenic sample. Input blot is from …
Figure 2 with 6 supplements
Active and repressed FLC-Venus transcriptional states are mitotically inherited.
(A) Unspliced FLC-Venus RNA in roots and shoots measured by qRT-PCR for non-vernalized (NV) plants or after 2, 4, 6, 8, or 10 weeks of cold followed by 7 days of warm. Data shown are mean ± s.e.m. …
Figure 2—figure supplement 1
FLC-Venus is quantitatively epigenetically repressed by cold exposure.
(A) FLC-Venus mRNA levels measured by RT-qPCR using primers specific for FLC-Venus. Data are normalized to UBC. 4WT0 indicates that plants were harvested immediately after exposure to 4 weeks of …
Figure 2—figure supplement 2
COOLAIR expression in transgenic FLC lines.
(A) Schematic of FLC genomic DNA with FLC mRNA and COOLAIR exons represented by black boxes. Dashed lines indicate splicing pattern. Coloured bars beneath COOLAIR transcripts represent the position …
Figure 2—figure supplement 3
Confocal microscopy of FLC-Venus and FLC-mCherry.
(A) Confocal microscopy images of non-vernalized FLC-Venus. FLC-Venus channel is a maximum intensity projection over 2–3 z-planes. Propidium iodide (red) was used to stain the cell wall; a single …
Figure 2—figure supplement 4
Confocal microscopy of FLC-Venus after vernalization.
FLC-Venus (yellow) in root meristems of plants exposed to 2, 4, 6, 8, or 10 weeks cold. Plants were imaged 7 days after return to warm. FLC-Venus channel is a maximum intensity projection over 2–3 …
Figure 2—figure supplement 5
Quantitative image analysis.
(A) Flowchart showing the processing steps used to calculate mean FLC-Venus intensity per cell. Cell wall (propidium iodide) images from each layer of the z-stack were masked and segmented. Each of …
Figure 2—figure supplement 6
Root-to-root variability in quantified images.
Four roots of a single FLC-Venus line exposed to 8 weeks of cold followed by 7 days of growth in warm. Propidium iodide, FLC-Venus, and Overlay are single z-plane images. ‘Quantified cells’ is the …
Figure 3 with 2 supplements
‘Mixed’ transcriptional states are mitotically inherited.
(A) In a trans memory system, the two copies of FLC are coordinately regulated and only two mitotically heritable states are possible (FLC-Venus/FLC-mCherry ON/ON, OFF/OFF). (B) In a cis memory …
Figure 3—figure supplement 1
Confocal images of NV FLC-Venus × FLC-mCherry F1 plants.
Confocal microscope images acquired simultaneously using dual excitation of FLC-Venus and FLC-mCherry. Images are maximum intensity projections over 2–4 z-planes. A single bright-field image …
Figure 3—figure supplement 2
Confocal images of vernalized FLC-Venus × FLC-mCherry F1 plants.
Confocal microscope images acquired simultaneously using dual excitation of FLC-Venus and FLC-mCherry. Images are maximum intensity projections over 2–4 z-planes. A single bright-field image …
