Imaging files for: ACD15, ACD21, and SLN regulate accumulation and mobility of MBD6 to silence genes and transposable elements

It's well known that DNA methylation is linked to gene silencing but
the mechanisms of how proteins that bind the DNA methylation cause gene
silencing remains unclear. We demonstrated that the novel MBD5/6 complex
contains three chaperone proteins, called ACD15, ACD21, and SLN, which
specifically mediate the gene silencing function. ACD15 and ACD21 bridge
the interaction of SLN to MBD5 and/or MBD6 while also functioning to drive
the accumulation of the MBD5/6 complex at CG methylation sites. We further
discovered that SLN also regulates the accumulation of the MBD5/6 complex
and regulates the turnover of all protein members once accumulated at meCG
sites.  To demonstrate these results we primarily used
fluorescence, confocal microscopy using RFP tagged MBD6, YFP tagged ACD15,
and CFP tagg ACD21 and SLN imaging the roots of Arabidopsis thaliana. We
expressed these constructs either alone or together in multiple mutant
lines including mbd5 mbd6, acd15, acd21, acd15 acd21, sln, and acd15 acd21
sln mutant plants. We also truncated MBD6 to remove the C terminus, called
the C term deletion, as well as a version of MBD6 lacking the C terminus
but with the domain necessary for interaction with ACD15 added back (MBD6
plus StkyC). With these microscopy experiments, we were able to
demonstrate the specificity of ACD15 for the StkyC domain, the function of
the chaperone proteins, and linke protein accumulation with gene
silencing.