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. 2015 Jan 6;5:760. doi: 10.3389/fpls.2014.00760

Figure 4.

Figure 4

SIHDACs interacted with MDAS-box proteins, TM29 and TAG1. (A) SIHDACs interacted with TM29 and TAG1 in yeast two-hybrid assays. SlHDA1, SlHDA3, and SlHDA4 were cloned into pGADT7 vector, whereas TM29 and TAG1 were cloned into pGBKT7 vector, respectively. The plasmids were cotransformed into the yeast strain AH109. The transformants were grown on the selective minimal medium without Leu and Trp (LW-) or without Leu, Trp, Ade and His (LWHA-). SlHDACs interacted with TM29 and TAG1 in Arabidopsis protoplasts in BiFC assays. (B) SlHDA1/SlHDA2/SlHDA4 fused with the N terminus (YN) and TAG1/TM29 fused with the C terminus (YC) of YFP were cotransformed into protoplasts and then incubated in the 100 μmol.m-2.s-1 for 12 h. The fluorescence was determined using a confocal microscope. The YFP fluorescence was excited by a 514 nm laser and captured at 523–600 nm, and the chlorophyll autofluorescence was captured at 650–750 nm. Bar = 20 μm. SlHDACs interacted with TM29 and TAG1 in pull-down assays. (C) GST- SlHDAC1, GST- SlHDAC3, GST- SlHDAC4 or GST was incubated with either TM29-His or TAG1-His and GST affinity resin, and the bound proteins were then eluted from resin and probed with the anti-His antibody.