Figure 5.

HY5 and SPL7 Coordinately Mediate MIR408 Expression in Response to Changing Light and Copper Conditions.

(A) RNA gel blot analysis of miR408 levels in wild-type, hy5, and spl7 seedlings in four different light and copper regimens. U6 small nuclear RNA was used as the loading control.

(B) and (C) RT-qPCR analysis of miR408 levels (B) and the miR408 target gene LAC13 (C) in wild-type, hy5, and spl7 seedlings. For comparison, levels of miR408 and LAC13 in DC/HL were set as 1.

(D) GUS activity in various transgenic plants. The pMIR408:GUS or pMIR408m:GUS reporter was expressed in the wild type or the indicated mutant background. Seedlings grown under different combinations of light and copper conditions were stained for GUS activity and visualized. Bars = 1 cm.

(E) RT-qPCR analysis of miR408 and pri-miR408 (top) and RNA gel blot analysis of miR408 levels (bottom) in wild-type, spl7, hy5, and hy5 spl7 seedlings under the DC/HL condition.

(F) RT-qPCR analysis of LAC13 transcript levels in the indicated genotypes under the DC/HL condition. Levels of miR408 and LAC13 in the wild type were set as 1.

Data for RT-qPCR are means ± sd (n = 3). The same letters indicate no statistical difference, while different letters denote groups with significant differences (ANOVA, P < 0.01).

(G) GUS activity in transgenic plants expressing pMIR408:GUS in the wild-type, spl7, hy5, and hy5 spl7 backgrounds. Bar = 1 cm.