Fig. 3

Fusion of the VP16 activator domain to UVR8 does not affect expression of suggested target genes. a, b, c, d, e, f The effect of fusing the VP16 activator domain to HY5 (VP16-HY5: a, d), UVR8 (VP16-UVR8: b, e) and HFR1 (VP16-HFR1: c, f) as measured by expression changes of target and negative control genes. Mutant protoplasts prepared from hy5-ks50, uvr8-7 and hfr1-101 seedlings were transfected with Pro _35S :3xHA-VP16AD-HY5/Pro _35S :HY5, Pro _35S :3xHA-VP16AD-UVR8/Pro _35S :UVR8 and Pro _35S :3xHA-VP16AD-HFR1/Pro _35S :HFR1 respectively. Gene expression was analysed 19–24 h post-transfection with protoplasts maintained under weak white light (a, b, c) and weak white light plus a final 3 h UV-B irradiation (d, e, f). Gene expression levels were normalized with the expression levels of the respective transfected effectors (i.e., VP16-HY5, HY5, VP16-UVR8, UVR8, VP16-HFR1, HFR1). Data is presented as relative expression in the presence of the effectors fused to VP16 to the respective control effectors without the VP16 domain (VP16-effector/effector). nd = non-determinable. Data are shown as the means of four (a, b, c) or three (d, e, f) biological replicates ± standard errors