FIGURE 3.

Copper deficiency does not impact the phenotype of cgl78-ami strains. A, CYC6 and CGL78 mRNA abundance was estimated by qRT-PCR. RNA was isolated from cells that were grown in medium containing nitrate to induce the amiRNA targeting CGL78, the presence of Cu(II)-EDTA in the medium is indicated. Control strains (black) or cgl78-ami lines (white) were grown in TAP medium to a density of 5–8 × 10⁶ cells/ml. Each symbol represents an independent experiment analyzed in technical duplicates. B, chlorophyll content in cells grown in medium supplemented either with ammonia or nitrate as nitrogen source and supplemented with or without copper as indicted. Error bars represent S.D. of at least three experiments. C, soluble fractions corresponding to 10 μg of protein were separated on sodium dodecyl sulfate-containing denaturing gels (15% acrylamide) and immuno-detection with anti-Cyt c₆ or OEE1 as a loading control. D, PSI/II-LHC super-complex formation is also diminished in copper-depleted cgl78-ami lines. Blue native PAGE of thylakoid membranes from CGL78 or cgl78-ami lines grown either in copper-replete or copper-depleted TAP medium with nitrate as nitrogen source. Experiments were performed at least twice.