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. 2020 Sep 2;184(3):1585–1600. doi: 10.1104/pp.20.00604

Figure 6.

Figure 6.

HDA15 can be phosphorylated in Arabidopsis etiolated seedlings. A, Sequence alignment of the HDA15 phosphorylation sites S448 and S452 with human and Arabidopsis HDACs. B, Immunoblotting of HDA15-GFP with the phospho-Ser antibody. HDA15-GFP immunoprecipitated from PromoterHDA15:HDA15-GFP/hda15-1 transgenic plants in 2-d-old etiolated seedlings (E) and 10-d-old seedlings grown under WL were analyzed by the phospho-Ser antibody after CIP treatment. GFP immunoprecipitated from PromoterHDA15:GFP/hda15-1 transgenic plants in 2-d-old E was used as a negative control. Star indicates the phosphorylated HDA15-GFP. Triangle indicates the phosphatase CIP. Immunoblotting with the GFP antibody indicates the input protein loading. C, Phos-tag gels of GFP-HDA15ZFHD. GFP-HDA15ZFHD immunoprecipitated from 35S:GFP-HDA15ZFHD/hda15-1 transgenic plants in 2-d-old E and 10-d-old seedlings grown under WL were analyzed by the Phos-tag gels or SDS-PAGE after CIP treatment. The asterisks in the Phos-tag gel indicate the phosphorylated GFP-HDA15ZFHD. CIP treatment resulted in GFP-HDA15ZFHD dephosphorylation (the band shifted down in the Phos-tag gels). WB, Western blot.